LOCUS       sequence-146169-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse cofilin S3A (nonphosphorylatable form)
            fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  AUTHORS   Chai X, Zhao S, Fan L, Zhang W, Lu X, Shao H, Wang S, Song L, Failla
            AV, Zobiak B, Mannherz HG, Frotscher M
  TITLE     Reelin and cofilin cooperate during the migration of cortical 
            neurons: a quantitative morphological analysis.
  JOURNAL   Development. 2016 Mar 15;143(6):1029-40. doi: 10.1242/dev.134163. 
            Epub 2016 Feb 18.
  PUBMED    26893343
REFERENCE   2  (bases 1 to 6404)
  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: 
            "10.1242/dev.134163"; journalName: "Development"; date: "2016-03-15-
            15"; volume: "143"; issue: "6"; pages: "1029-40"
COMMENT     Sequence Label: pCAG-cofilin-S3A
FEATURES             Location/Qualifiers
     source          1..6404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        218..521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..800
                     /label=chicken beta-actin promoter
     intron          802..1819
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1743..1765
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1827..1846
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1879..1888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2417..2426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2423..2425,2426..2428,2429..3142)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2423..2425,2426..2428,2429..3139)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2468..2489)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2729..2748)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3076..3097
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3151..3627
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3350..3369)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3667..3807
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3667..3796
                     /label=AAV2 ITR
     rep_origin      3882..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3969..3988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4179..4200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4354..4373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4473..4495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4533..4551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4619..4723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4724..4792,4793..5584)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4942..4961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5755..6343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6244..6263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtactag ttattaatag taatcaatta
      181 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      241 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      301 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      361 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      421 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      481 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      541 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      601 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
      661 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      721 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      781 agcgaagcgc gcggcgggcg gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
      841 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      901 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
      961 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     1021 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     1081 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     1141 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     1201 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     1261 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     1321 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     1381 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     1441 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     1501 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     1561 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     1621 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     1681 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     1741 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     1801 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1861 tggcaaagaa ttgaattcgc caccatggcc gctggtgtgg ctgtctctga tggtgtcatc
     1921 aaggtgttca atgacatgaa ggttcgcaag tcttcaacac cagaagaagt gaagaaacgc
     1981 aagaaggcgg tgctcttttg cctgagtgag gacaagaaga acatcatcct ggaggagggc
     2041 aaggagatcc tggtaggaga tgtggggcag actgtggacg acccctacac cacttttgtc
     2101 aagatgctgc cagacaagga ctgccgctat gcactctatg atgcaaccta tgagaccaag
     2161 gagagcaaga aggaggacct ggtgttcatc ttctgggccc ccgagaatgc acccctcaag
     2221 agcaaaatga tctatgccag ctccaaggat gccatcaaga agaagctgac aggaatcaag
     2281 catgaattac aagctaactg ctacgaggag gtcaaggacc gctgcaccct ggcagagaaa
     2341 ctaggtggca gcgccgtcat ttccctggag ggcaagcctt tgaagcttgc ctcgagcagc
     2401 gctgctcgag agatctgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2461 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2521 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2581 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2641 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2701 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2761 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2821 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2881 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2941 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3001 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3061 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3121 catggacgag ctgtacaagt aaagatctac gggtggcatc cctgtgaccc ctccccagtg
     3181 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3241 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3301 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3361 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3421 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3481 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3541 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3601 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3661 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3721 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3781 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3841 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4321 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4381 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4441 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4501 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4561 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5881 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
