LOCUS       sequence-490232-        9652 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Silencing of endogenous cofilin in rat/mouse neurons with replacment
            by neuronal specific expression of K22Q mutant of human cofilin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9652)
  TITLE     Bamburg lab plasmids 2025
REFERENCE   2  (bases 1 to 9652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NSE-huCof K22Q + rat cof shRNA
FEATURES             Location/Qualifiers
     source          1..9652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..153
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     241..391
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     CDS             2142..2819
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2142..2819
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2272..2290)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2540..2559
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2734..2757
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2979..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3070..3089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3165..3379
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     3300..3319
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     repeat_region   6661..6763
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6852..6869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7023..7611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7103..7122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             8443..9237
                     /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             8443..9237
                     /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(8497..8516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9107..9126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(join(9285..9652,1..13))
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9347..9368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9559..9578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctacagggcg cgtccattcg ccattcagga tcgaattaat tcttaattaa catcatcaat
       61 aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt ttgtgacgtg
      121 gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt gatgttgcaa
      181 gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg gtgtgcgccg
      241 gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag taaatttggg
      301 cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga agtgaaatct
      361 gaataatttt gtgttactca tagcgcgtaa tactgtaatc ttaaggggac agtaaaggtg
      421 atggcaggaa ggcagccccc ggaggtcaaa ggctgggcac gcgggaggag aggccagagt
      481 cagaggctgc gggtatctca gatatgaagg aaagatgaga gaggctcagg aagaggtaag
      541 aaaagacaca agagaccaga gaagggagaa gaattagaga gggaggcaga ggaccgctgt
      601 ctctacagac atagctggta gagactggga ggaagggatg aaccctgagc gcatgaaggg
      661 aaggaggtgg ctggtggtat atggaggatg tagctgggcc agggaaaaga tcctgcacta
      721 aaaatctgaa gctaaaaata acaggacacg gggtggagag gcgaaaggag ggcagattga
      781 ggcagagaga ctgagaggcc tggggatgtg ggcattccgg tagggcacac agttcacttg
      841 tcttctcttt ttccaggagg ccaaagatgc tgacctcaag aactcataat accccagtgg
      901 ggaccaccgc attcatagcc ctgttacaag aagtgggaga tgttcctttt tgtcccagac
      961 tggaaatcca ttacatcccg aggctcaggt tctgtggtgg tcatctctgt gtggcttgtt
     1021 ctgtgggcct acctaaagtc ctaagcacag ctctcaagca gatccgaggc gactaagatg
     1081 ctagtagggg ttgtctggag agaagagccg aggaggtggg ctgtgatgga tcagttcagc
     1141 tttcaaataa aaaggcgttt ttatattctg tgtcgagttc gtgaacccct gtggtgggct
     1201 tctccatctg tctgggttag tacctgccac tatactggaa taaggggacg cctgcttccc
     1261 tcgagttggc tggacaaggt tatgagcatc cgtgtactta tggggttgcc agcttggtcc
     1321 tggatcgccc gggcccttcc cccacccgtt cggttcccca ccaccacccg cgctcgtacg
     1381 tgcgtctccg cctgcagctc ttgactcatc ggggcccccg ggtcacatgc gctcgctcgg
     1441 ctctataggc gccgccccct gcccaccccc cgcccgcgct gggagccgca gccgccgcca
     1501 ctcctgctct ctctgcgccg ccgccgtcac caccgccacc gccaccggct gagtctgcag
     1561 tcctccgaaa cgggccctct agactcgagc ggccgctcga gcctaagctt gatatcgaat
     1621 tcaccaccat ggcctccggt gtggctgtct ctgatggtgt catcaaggtg ttcaacgaca
     1681 tgaaggtgcg tcagagctca accccagagg aggtgaagaa gcgcaagaag gcggtgctct
     1741 tctgcctgag tgaggacaag aagaacatca tcctggagga gggcaaggag atcctggtgg
     1801 gcgatgtggg ccagactgtc gacgatccct acgccacctt tgtcaagatg ctgccagata
     1861 aggactgccg ctatgccctc tatgatgcaa cctatgagac caaggagagc aagaaggagg
     1921 atctggtgtt tatcttctgg gcccccgagt ctgcgcccct taagagcaaa atgatttatg
     1981 ccagctccaa ggacgccatc aagaagaagc tgacagggat caagcatgaa ttgcaagcaa
     2041 actgctacga ggaggtcaag gaccgctgca ccctggcaga gaagctgggg ggcagtgcgg
     2101 tcatctccct ggagggcaag cctttgttat cccgggatcc gatggcctcc tccgaggacg
     2161 tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt
     2221 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2281 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct cagttccagt
     2341 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2401 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2461 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2521 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     2581 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagatg aggctgaagc
     2641 tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacatggcc aagaagcccg
     2701 tgcagctgcc cggcgcctac aagaccgaca tcaagctgga catcacctcc cacaacgagg
     2761 actacaccat cgtggaacag tacgagcgcg ccgagggccg ccactccacc ggcgcctaag
     2821 aattggccgc gactctagtg gatccgagct cggtaccaag cttaagtttc tagatcataa
     2881 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2941 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3061 attctagttg tggtttgtcc aaactcatca atgtatctta atcgactcga agatccacta
     3121 gtaacggccg ccagtgtgct ggaattcgcc cttccatgga attcgaacgc tgacgtcatc
     3181 aacccgctcc aaggaatcgc gggcccagtg tcactaggcg ggaacaccca gcgcgcgtgc
     3241 gccctggcag gaagatggct gtgagggaca ggggagtggc gccctgcaat atttgcatgt
     3301 cgctatgtgt tctgggaaat caccataaac gtgaaatgtc tttggatttg ggaatcttat
     3361 aagttctgta tgagaccaca gatctaaggt gttcaatgac atgaaattct cgagatttca
     3421 tgtcattgaa cacctttttt tggaaagctt gatatcgaat tcctgcagcc cgggggatct
     3481 gggcgtggtt aagggtggga aagaatatat aaggtggggg tcttatgtag ttttgtatct
     3541 gttttgcagc agccgccgcc gccatgagca ccaactcgtt tgatggaagc attgtgagct
     3601 catatttgac aacgcgcatg cccccatggg ccggggtgcg tcagaatgtg atgggctcca
     3661 gcattgatgg tcgccccgtc ctgcccgcaa actctactac cttgacctac gagaccgtgt
     3721 ctggaacgcc gttggagact gcagcctccg ccgccgcttc agccgctgca gccaccgccc
     3781 gcgggattgt gactgacttt gctttcctga gcccgcttgc aagcagtgca gcttcccgtt
     3841 catccgcccg cgatgacaag ttgacggctc ttttggcaca attggattct ttgacccggg
     3901 aacttaatgt cgtttctcag cagctgttgg atctgcgcca gcaggtttct gccctgaagg
     3961 cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa accagactct gtttggattt
     4021 ggatcaagca agtgtcttgc tgtctttatt taggggtttt gcgcgcgcgg taggcccggg
     4081 accagcggtc tcggtcgttg agggtcctgt gtattttttc caggacgtgg taaaggtgac
     4141 tctggatgtt cagatacatg ggcataagcc cgtctctggg gtggaggtag caccactgca
     4201 gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc gtagcaggag cgctgggcgt
     4261 ggtgcctaaa aatgtctttc agtagcaagc tgattgccag gggcaggccc ttggtgtaag
     4321 tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg ggatatgaga tgcatcttgg
     4381 actgtatttt taggttggct atgttcccag ccatatccct ccggggattc atgttgtgca
     4441 gaaccaccag cacagtgtat ccggtgcact tgggaaattt gtcatgtagc ttagaaggaa
     4501 atgcgtggaa gaacttggag acgcccttgt gacctccaag attttccatg cattcgtcca
     4561 taatgatggc aatgggccca cgggcggcgg cctgggcgaa gatatttctg ggatcactaa
     4621 cgtcatagtt gtgttccagg atgagatcgt cataggccat ttttacaaag cgcgggcgga
     4681 gggtgccaga ctgcggtata atggttccat ccggcccagg ggcgtagtta ccctcacaga
     4741 tttgcatttc ccacgctttg agttcagatg gggggatcat gtctacctgc ggggcgatga
     4801 agaaaacggt ttccggggta ggggagatca gctgggaaga aagcaggttc ctgagcagct
     4861 gcgacttacc gcagccggtg ggcccgtaaa tcacacctat taccggctgc aactggtagt
     4921 taagagagct gcagctgccg tcatccctga gcaggggggc cacttcgtta agcatgtccc
     4981 tgactcgcat gttttccctg accaaatccg ccagaaggcg ctcgccgccc agcgatagca
     5041 gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc gtccgccgta ggcatgcttt
     5101 tgagcgtttg accaagcagt tccaggcggt cccacagctc ggtcacctgc tctacggcat
     5161 ctcgatccag catatctcct cgtttcgcgg gttggggcgg ctttcgctgt acggcagtag
     5221 tcggtgctcg tccagacggg ccagggtcat gtctttccac gggcgcaggg tcctcgtcag
     5281 cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc gcgctggcca gggtgcgctt
     5341 gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg ccctgcgcgt cggccaggta
     5401 gcatttgacc atggtgtcat agtccagccc ctccgcggcg tggcccttgg cgcgcagctt
     5461 gcccttggag gaggcgccgc acgaggggca gtgcagactt ttgagggcgt agagcttggg
     5521 cgcgagaaat accgattccg gggagtaggc atccgcgccg caggccccgc agacggtctc
     5581 gcattccacg agccaggtga gctctggccg ttcggggtca aaaaccaggt ttcccccatg
     5641 ctttttgatg cgtttcttac ctctggtttc catgagccgg tgtccacgct cggtgacgaa
     5701 aaggctgtcc gtgtccccgt atacagactt gagagggagt ttaaacgaat tcaatagctt
     5761 gttgcatggg cggcgatata aaatgcaagg tgctgctcaa aaaatcaggc aaagcctcgc
     5821 gcaaaaaaga aagcacatcg tagtcatgct catgcagata aaggcaggta agctccggaa
     5881 ccaccacaga aaaagacacc atttttctct caaacatgtc tgcgggtttc tgcataaaca
     5941 caaaataaaa taacaaaaaa acatttaaac attagaagcc tgtcttacaa caggaaaaac
     6001 aacccttata agcataagac ggactacggc catgccggcg tgaccgtaaa aaaactggtc
     6061 accgtgatta aaaagcacca ccgacagctc ctcggtcatg tccggagtca taatgtaaga
     6121 ctcggtaaac acatcaggtt gattcacatc ggtcagtgct aaaaagcgac cgaaatagcc
     6181 cgggggaata catacccgca ggcgtagaga caacattaca gcccccatag gaggtataac
     6241 aaaattaata ggagagaaaa acacataaac acctgaaaaa ccctcctgcc taggcaaaat
     6301 agcaccctcc cgctccagaa caacatacag cgcttccaca gcggcagcca taacagtcag
     6361 ccttaccagt aaaaaagaaa acctattaaa aaaacaccac tcgacacggc accagctcaa
     6421 tcagtcacag tgtaaaaaag ggccaagtgc agagcgagta tatataggac taaaaaatga
     6481 cgtaacggtt aaagtccaca aaaaacaccc agaaaaccgc acgcgaacct acgcccagaa
     6541 acgaaagcca aaaaacccac aacttcctca aatcgtcact tccgttttcc cacgttacgt
     6601 cacttcccat tttaagaaaa ctacaattcc caacacatac aagttactcc gccctaaaac
     6661 ctacgtcacc cgccccgttc ccacgccccg cgccacgtca caaactccac cccctcatta
     6721 tcatattggc ttcaatccaa aataaggtat attattgatg atgttaatta acatgcatgg
     6781 atccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6841 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6901 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6961 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7021 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7081 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7141 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7201 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7261 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7321 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7381 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7441 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7501 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7741 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     8101 gccatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa agccagtccg
     8161 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     8221 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     8281 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     8341 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     8401 ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     8461 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     8521 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     8581 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     8641 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     8701 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     8761 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     8821 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     8881 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     8941 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     9001 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     9061 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     9121 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     9181 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     9241 ttgttaaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     9301 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     9361 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     9421 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     9481 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     9541 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     9601 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cg
//
