LOCUS       sequence-146170-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse cofilin S3D fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  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 6406)
  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-S3D
FEATURES             Location/Qualifiers
     source          1..6406
                     /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      2419..2428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2425..2427,2428..2430,2431..3144)
                     /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(2425..2427,2428..2430,2431..3141)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2470..2491)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2731..2750)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3078..3099
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3153..3629
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3352..3371)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3669..3809
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3669..3798
                     /label=AAV2 ITR
     rep_origin      3884..4339
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3971..3990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4181..4202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4356..4375)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4475..4497
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4535..4553)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4621..4725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4726..4794,4795..5586)
                     /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(4944..4963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /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 gatggtgtgg 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 tgcgaagctt gcctcgagca
     2401 gcgctgctcg agagatctgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2461 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2521 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2581 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2641 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2701 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2761 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2821 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2881 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2941 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3001 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3061 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3121 ggcatggacg agctgtacaa gtaaagatct acgggtggca tccctgtgac ccctccccag
     3181 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3241 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3301 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3361 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3421 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3481 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3541 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3601 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3661 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3721 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3781 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3841 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3901 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3961 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4021 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4081 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4141 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4201 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4261 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4321 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4381 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4441 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4501 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4561 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
