LOCUS       sequence-375581-        5292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  F-actin structures labelling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5292)
  AUTHORS   Alfadil E, Bradke F, Dupraz S
  TITLE     In Situ Visualization of Axon Growth and Growth Cone Dynamics in 
            Acute Ex Vivo Embryonic Brain Slice Cultures.
  JOURNAL   J Vis Exp. 2021 Oct 14;(176). doi: 10.3791/63068.
  PUBMED    34723948
REFERENCE   2  (bases 1 to 5292)
  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"; journalName: "J Vis 
            Exp."; date: "2021-10-14"; volume: "(176). doi"; pages: " 
            10.3791/63068"
COMMENT     Sequence Label: pTub-alpha1-Lifeact-GFP
FEATURES             Location/Qualifiers
     source          1..5292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1166..1216
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      1232..1241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1238..1240,1241..1243,1244..1957)
                     /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(1238..1240,1241..1243,1244..1954)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1283..1304)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1544..1563)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1891..1912
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2080..2201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2117..2136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2171..2190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2208..2663)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2345..2366)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2557..2576
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2690..2794
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2796..3153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2796..2987
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2819..2839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3004..3139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3188..3982
                     /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             3188..3982
                     /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(3242..3261)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3852..3871
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4170..4189)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4214..4261
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4590..5178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5079..5098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattat gctgaattcc gtattaggag ggatggctca tttctaggga caaaataaca
       61 caggctctgg ggtgaggtag ggtcgggtaa gggggcgagc ggggaggggt gatgggagag
      121 cactaccgct aaagaaagca agctctctag taaagcgtta agtactactt tgtatattgt
      181 ttctcttttt tcctttctct ctctctctct ctttgtttgt ttttggtttt tgagacagga
      241 tatcacacta tatatccagg ccgggtacaa aataaccgca gtcctccgct tcccaatgct
      301 gggatttaca ggcataaacc actaagggcg gttatgatga ccttgagctg caagtcttcc
      361 tgcctctgcc tcccgggtgc tgttgagggt cataggcgtg gtctattcat actgagcttc
      421 tgaattttgc tcaaatatta ataataatag taataataat aaccacaata atacaactgt
      481 aaaactaaac atttacccac gcctttttga ccatcattcc catagctctt gctactttat
      541 ttaaagcgaa cagagatgtt gaatcctgac ggaacgtatt taaatttagt gtagtataaa
      601 tgaaaagctg gaatttacca taaagaatct caacacaaat tctgtgatta agtgttgggg
      661 aaaaccttaa attatcctaa ctacagttta aggtttaagc tccgtataat cacccaaccc
      721 ccgttttctt tcttccctct ctacccctcc ccagctccac cccataatgg atgctcggct
      781 agttgctttt gcgaggcctt tgtctgaagg atgcaaaatc tacggatgct agcgaggggg
      841 ggaagggggg agagattacc tcataccatg tcgcttgcac caatcaccac tctcctgtcg
      901 cggcttctct gggcagacgg aggggtctgg accaacagga aaaggccttg gcccatcccc
      961 atggtgaccg agctgtatat aaggaggcgc atccgcccaa gtgccgcagg ttctcttaca
     1021 tcgaccgctt aagagtcgcg ctgtaagaag caacacctcc tcctcgcctc cgccatccac
     1081 ccggcagccg cgaagcagca acaccggtga gcgggggatc cactagcgct accggactca
     1141 gatctcgagc tcaagcttcg aattcatggg tgtcgcagat ttgatcaaga aattcgaaag
     1201 catctcaaag gaagaagggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct
     1261 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1321 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1381 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1441 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1501 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1561 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1621 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1681 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1741 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1801 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1861 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1921 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     1981 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2041 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2101 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2161 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2221 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2281 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2341 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2401 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2461 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2521 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2581 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2641 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2701 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2761 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2821 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3061 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3121 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3181 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3241 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3301 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3361 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3421 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3481 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3541 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3601 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3661 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3721 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3781 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3841 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3901 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3961 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4021 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4081 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4141 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4201 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4261 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4321 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4381 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4441 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4501 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4561 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4681 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4741 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4801 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4861 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4921 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4981 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5041 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5101 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5161 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5221 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5281 accgccatgc at
//
