LOCUS       sequence-345857-        5265 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 5265)
  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 5265)
  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-iLifeact-NeonGreen
FEATURES             Location/Qualifiers
     source          1..5265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..837
                     /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             43..837
                     /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(97..116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     707..726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1025..1044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1069..1116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1445..2033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1934..1953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3293..3343
                     /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"
     CDS             3368..4075
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      3620..3629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4200..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4237..4256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4291..4310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4328..4783)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4465..4486)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4677..4696
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4810..4914
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4916..5265,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4916..5107
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4939..4959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5124..5259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
       61 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      121 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
      181 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
      241 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
      301 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
      361 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
      421 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
      481 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
      541 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
      601 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
      661 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
      721 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
      781 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
      841 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
      901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
      961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     1021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     1141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     1201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     1261 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     1321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2101 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattatgct
     2161 gaattccgta ttaggaggga tggctcattt ctagggacaa aataacacag gctctggggt
     2221 gaggtagggt cgggtaaggg ggcgagcggg gaggggtgat gggagagcac taccgctaaa
     2281 gaaagcaagc tctctagtaa agcgttaagt actactttgt atattgtttc tcttttttcc
     2341 tttctctctc tctctctctt tgtttgtttt tggtttttga gacaggatat cacactatat
     2401 atccaggccg ggtacaaaat aaccgcagtc ctccgcttcc caatgctggg atttacaggc
     2461 ataaaccact aagggcggtt atgatgacct tgagctgcaa gtcttcctgc ctctgcctcc
     2521 cgggtgctgt tgagggtcat aggcgtggtc tattcatact gagcttctga attttgctca
     2581 aatattaata ataatagtaa taataataac cacaataata caactgtaaa actaaacatt
     2641 tacccacgcc tttttgacca tcattcccat agctcttgct actttattta aagcgaacag
     2701 agatgttgaa tcctgacgga acgtatttaa atttagtgta gtataaatga aaagctggaa
     2761 tttaccataa agaatctcaa cacaaattct gtgattaagt gttggggaaa accttaaatt
     2821 atcctaacta cagtttaagg tttaagctcc gtataatcac ccaacccccg ttttctttct
     2881 tccctctcta cccctcccca gctccacccc ataatggatg ctcggctagt tgcttttgcg
     2941 aggcctttgt ctgaaggatg caaaatctac ggatgctagc gaggggggga aggggggaga
     3001 gattacctca taccatgtcg cttgcaccaa tcaccactct cctgtcgcgg cttctctggg
     3061 cagacggagg ggtctggacc aacaggaaaa ggccttggcc catccccatg gtgaccgagc
     3121 tgtatataag gaggcgcatc cgcccaagtg ccgcaggttc tcttacatcg accgcttaag
     3181 agtcgcgctg taagaagcaa cacctcctcc tcgcctccgc catccacccg gcagccgcga
     3241 agcagcaaca ccggtgagcg ggggatctcg agctcaagct tcgaattcca ccatgggcgt
     3301 ggccgacctg atcaagaagt tcgagagcat cagcaaggag gagggggatc caccggtcgc
     3361 caccctggtg agcaagggcg aggaggataa catggcctct ctcccagcga cacatgagtt
     3421 acacatcttt ggctccatca acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa
     3481 tccaaatgat ggttatgagg agttaaacct gaagtccacc aagggtgacc tccagttctc
     3541 cccctggatt ctggtccctc atatcgggta tggcttccat cagtacctgc cctaccctga
     3601 cgggatgtcg cctttccagg ccgccatggt agatggctcc ggctaccaag tccatcgcac
     3661 aatgcagttt gaagatggtg cctcccttac tgttaactac cgctacacct acgagggaag
     3721 ccacatcaaa ggagaggccc aggtgaaggg gactggtttc cctgctgacg gtcctgtgat
     3781 gaccaactcg ctgaccgctg cggactggtg caggtcgaag aagacttacc ccaacgacaa
     3841 aaccatcatc agtaccttta agtggagtta caccactgga aatggcaagc gctaccggag
     3901 cactgcgcgg accacctaca cctttgccaa gccaatggcg gctaactatc tgaagaacca
     3961 gccgatgtac gtgttccgta agacggagct caagcactcc aagaccgagc tcaacttcaa
     4021 ggagtggcaa aaggccttta ccgatgtgat gggcatggac gagctgtaca agtaatgagc
     4081 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4141 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4321 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4381 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4441 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4501 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4561 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4621 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4681 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4741 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4801 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4861 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4921 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4981 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5041 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5101 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5161 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5221 agctattcca gaagtagtga ggaggctttt ttggaggcct aggct
//
