LOCUS       sequence-490155-        8671 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Expression of cof with two of 4 cys mutated to A. Does NOT 
            incorporate into cofilin-actin rods ..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8671)
  AUTHORS   Bernstein BW, Shaw AE, Minamide LS, Pak CW, Bamburg JR
  TITLE     Incorporation of cofilin into rods depends on disulfide 
            intermolecular bonds: implications for actin regulation and 
            neurodegenerative disease.
  JOURNAL   J Neurosci. 2012 May 9;32(19):6670-81. doi: 
            10.1523/JNEUROSCI.6020-11.2012.
  PUBMED    22573689
REFERENCE   2  (bases 1 to 8671)
  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"; doi: 
            "10.1523/JNEUROSCI.6020-11"; journalName: "J Neurosci"; date: 
            "2012-05-9- 9"; volume: "32"; issue: "19"; pages: "6670-81"
COMMENT     Sequence Label: huCof C39A,C147A-mRFP
FEATURES             Location/Qualifiers
     source          1..8671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(22..402)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(84..105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     296..315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   440..542
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     630..780
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        838..1141
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1142..1345
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1296..1316
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1944..2621
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1944..2621
                     /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(2074..2092)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2342..2361
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2536..2559
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2753..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2844..2863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6065..6167
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6260..6277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6431..7019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6511..6530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7851..8645
                     /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             7851..8645
                     /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(7905..7924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8515..8534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
       61 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      121 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      181 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      241 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      301 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      361 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggat
      421 cgaattaatt cttaattaac atcatcaata atatacctta ttttggattg aagccaatat
      481 gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt
      541 agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt
      601 ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg
      661 ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg
      721 ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat
      781 actgtaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      841 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      901 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      961 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1021 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1081 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1141 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1201 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1261 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1321 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatctgg
     1381 taccgtcgac gcggccgctc gagcctaagc ttgatatcga attcaccacc atggcctccg
     1441 gtgtggctgt ctctgatggt gtcatcaagg tgttcaacga catgaaggtg cgtaagagct
     1501 caaccccaga ggaggtgaag aagcgcaaga aggcggtgct cttcgccctg agtgaggaca
     1561 agaagaacat catcctggag gagggcaagg agatcctggt gggcgatgtg ggccagactg
     1621 tcgacgatcc ctacgccacc tttgtcaaga tgctgccaga taaggactgc cgctatgccc
     1681 tctatgatgc aacctatgag accaaggaga gcaagaagga ggatctggtg tttatcttct
     1741 gggcccccga gtctgcgccc cttaagagca aaatgattta tgccagctcc aaggacgcca
     1801 tcaagaagaa gctgacaggg atcaagcatg aattgcaagc aaactgctac gaggaggtca
     1861 aggaccgcgc gaccctggca gagaagctgg ggggcagtgc ggtcatctcc ctggagggca
     1921 agcctttgtt atcccgggat ccgatggcct cctccgagga cgtcatcaag gagttcatgc
     1981 gcttcaaggt gcgcatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2041 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     2101 ccctgccctt cgcctgggac atcctgtccc ctcagttcca gtacggctcc aaggcctacg
     2161 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     2221 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     2281 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     2341 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgg atgtaccccg
     2401 aggacggcgc cctgaagggc gagatcaaga tgaggctgaa gctgaaggac ggcggccact
     2461 acgacgccga ggtcaagacc acctacatgg ccaagaagcc cgtgcagctg cccggcgcct
     2521 acaagaccga catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     2581 agtacgagcg cgccgagggc cgccactcca ccggcgccta agaattggcc gcgactctag
     2641 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2701 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgg
     2881 atctgggcgt ggttaagggt gggaaagaat atataaggtg ggggtcttat gtagttttgt
     2941 atctgttttg cagcagccgc cgccgccatg agcaccaact cgtttgatgg aagcattgtg
     3001 agctcatatt tgacaacgcg catgccccca tgggccgggg tgcgtcagaa tgtgatgggc
     3061 tccagcattg atggtcgccc cgtcctgccc gcaaactcta ctaccttgac ctacgagacc
     3121 gtgtctggaa cgccgttgga gactgcagcc tccgccgccg cttcagccgc tgcagccacc
     3181 gcccgcggga ttgtgactga ctttgctttc ctgagcccgc ttgcaagcag tgcagcttcc
     3241 cgttcatccg cccgcgatga caagttgacg gctcttttgg cacaattgga ttctttgacc
     3301 cgggaactta atgtcgtttc tcagcagctg ttggatctgc gccagcaggt ttctgccctg
     3361 aaggcttcct cccctcccaa tgcggtttaa aacataaata aaaaaccaga ctctgtttgg
     3421 atttggatca agcaagtgtc ttgctgtctt tatttagggg ttttgcgcgc gcggtaggcc
     3481 cgggaccagc ggtctcggtc gttgagggtc ctgtgtattt tttccaggac gtggtaaagg
     3541 tgactctgga tgttcagata catgggcata agcccgtctc tggggtggag gtagcaccac
     3601 tgcagagctt catgctgcgg ggtggtgttg tagatgatcc agtcgtagca ggagcgctgg
     3661 gcgtggtgcc taaaaatgtc tttcagtagc aagctgattg ccaggggcag gcccttggtg
     3721 taagtgttta caaagcggtt aagctgggat gggtgcatac gtggggatat gagatgcatc
     3781 ttggactgta tttttaggtt ggctatgttc ccagccatat ccctccgggg attcatgttg
     3841 tgcagaacca ccagcacagt gtatccggtg cacttgggaa atttgtcatg tagcttagaa
     3901 ggaaatgcgt ggaagaactt ggagacgccc ttgtgacctc caagattttc catgcattcg
     3961 tccataatga tggcaatggg cccacgggcg gcggcctggg cgaagatatt tctgggatca
     4021 ctaacgtcat agttgtgttc caggatgaga tcgtcatagg ccatttttac aaagcgcggg
     4081 cggagggtgc cagactgcgg tataatggtt ccatccggcc caggggcgta gttaccctca
     4141 cagatttgca tttcccacgc tttgagttca gatgggggga tcatgtctac ctgcggggcg
     4201 atgaagaaaa cggtttccgg ggtaggggag atcagctggg aagaaagcag gttcctgagc
     4261 agctgcgact taccgcagcc ggtgggcccg taaatcacac ctattaccgg ctgcaactgg
     4321 tagttaagag agctgcagct gccgtcatcc ctgagcaggg gggccacttc gttaagcatg
     4381 tccctgactc gcatgttttc cctgaccaaa tccgccagaa ggcgctcgcc gcccagcgat
     4441 agcagttctt gcaaggaagc aaagtttttc aacggtttga gaccgtccgc cgtaggcatg
     4501 cttttgagcg tttgaccaag cagttccagg cggtcccaca gctcggtcac ctgctctacg
     4561 gcatctcgat ccagcatatc tcctcgtttc gcgggttggg gcggctttcg ctgtacggca
     4621 gtagtcggtg ctcgtccaga cgggccaggg tcatgtcttt ccacgggcgc agggtcctcg
     4681 tcagcgtagt ctgggtcacg gtgaaggggt gcgctccggg ctgcgcgctg gccagggtgc
     4741 gcttgaggct ggtcctgctg gtgctgaagc gctgccggtc ttcgccctgc gcgtcggcca
     4801 ggtagcattt gaccatggtg tcatagtcca gcccctccgc ggcgtggccc ttggcgcgca
     4861 gcttgccctt ggaggaggcg ccgcacgagg ggcagtgcag acttttgagg gcgtagagct
     4921 tgggcgcgag aaataccgat tccggggagt aggcatccgc gccgcaggcc ccgcagacgg
     4981 tctcgcattc cacgagccag gtgagctctg gccgttcggg gtcaaaaacc aggtttcccc
     5041 catgcttttt gatgcgtttc ttacctctgg tttccatgag ccggtgtcca cgctcggtga
     5101 cgaaaaggct gtccgtgtcc ccgtatacag acttgagagg gagtttaaac gaattcaata
     5161 gcttgttgca tgggcggcga tataaaatgc aaggtgctgc tcaaaaaatc aggcaaagcc
     5221 tcgcgcaaaa aagaaagcac atcgtagtca tgctcatgca gataaaggca ggtaagctcc
     5281 ggaaccacca cagaaaaaga caccattttt ctctcaaaca tgtctgcggg tttctgcata
     5341 aacacaaaat aaaataacaa aaaaacattt aaacattaga agcctgtctt acaacaggaa
     5401 aaacaaccct tataagcata agacggacta cggccatgcc ggcgtgaccg taaaaaaact
     5461 ggtcaccgtg attaaaaagc accaccgaca gctcctcggt catgtccgga gtcataatgt
     5521 aagactcggt aaacacatca ggttgattca catcggtcag tgctaaaaag cgaccgaaat
     5581 agcccggggg aatacatacc cgcaggcgta gagacaacat tacagccccc ataggaggta
     5641 taacaaaatt aataggagag aaaaacacat aaacacctga aaaaccctcc tgcctaggca
     5701 aaatagcacc ctcccgctcc agaacaacat acagcgcttc cacagcggca gccataacag
     5761 tcagccttac cagtaaaaaa gaaaacctat taaaaaaaca ccactcgaca cggcaccagc
     5821 tcaatcagtc acagtgtaaa aaagggccaa gtgcagagcg agtatatata ggactaaaaa
     5881 atgacgtaac ggttaaagtc cacaaaaaac acccagaaaa ccgcacgcga acctacgccc
     5941 agaaacgaaa gccaaaaaac ccacaacttc ctcaaatcgt cacttccgtt ttcccacgtt
     6001 acgtcacttc ccattttaag aaaactacaa ttcccaacac atacaagtta ctccgcccta
     6061 aaacctacgt cacccgcccc gttcccacgc cccgcgccac gtcacaaact ccaccccctc
     6121 attatcatat tggcttcaat ccaaaataag gtatattatt gatgatgtta attaacatgc
     6181 atggatcgat ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6241 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6841 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7321 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7501 ttgctgcagc catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag
     7561 ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa
     7621 gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc
     7681 tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg
     7741 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
     7801 ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     7861 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7921 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7981 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     8041 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     8101 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     8161 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     8221 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     8281 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     8341 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     8401 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     8461 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     8521 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     8581 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     8641 tctgaatttt gttaaaattt ttgttaaatc a
//
