LOCUS       sequence-349045-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  labelling of microtubule plus end and F-actin structures.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  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 6064)
  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-alpha-EB3-NeonGreen
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        42..233
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(65..85)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      250..385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             434..1228
                     /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             434..1228
                     /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(488..507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1098..1117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1416..1435)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1460..1507
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1836..2424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2325..2344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3691..3700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4549..4558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4555..5265
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4810..4819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5390..5511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5427..5446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5481..5500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5518..5973)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5655..5676)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5867..5886
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6000..6064,1..40)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
       61 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
      121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      301 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
      421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      601 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      961 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     1021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     1081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     1141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     1201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     1261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     1321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1381 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     1441 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     1501 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     1561 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     1621 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     1681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2521 cgtattaccg ccatgcatta gttattatgc tgaattccgt attaggaggg atggctcatt
     2581 tctagggaca aaataacaca ggctctgggg tgaggtaggg tcgggtaagg gggcgagcgg
     2641 ggaggggtga tgggagagca ctaccgctaa agaaagcaag ctctctagta aagcgttaag
     2701 tactactttg tatattgttt ctcttttttc ctttctctct ctctctctct ttgtttgttt
     2761 ttggtttttg agacaggata tcacactata tatccaggcc gggtacaaaa taaccgcagt
     2821 cctccgcttc ccaatgctgg gatttacagg cataaaccac taagggcggt tatgatgacc
     2881 ttgagctgca agtcttcctg cctctgcctc ccgggtgctg ttgagggtca taggcgtggt
     2941 ctattcatac tgagcttctg aattttgctc aaatattaat aataatagta ataataataa
     3001 ccacaataat acaactgtaa aactaaacat ttacccacgc ctttttgacc atcattccca
     3061 tagctcttgc tactttattt aaagcgaaca gagatgttga atcctgacgg aacgtattta
     3121 aatttagtgt agtataaatg aaaagctgga atttaccata aagaatctca acacaaattc
     3181 tgtgattaag tgttggggaa aaccttaaat tatcctaact acagtttaag gtttaagctc
     3241 cgtataatca cccaaccccc gttttctttc ttccctctct acccctcccc agctccaccc
     3301 cataatggat gctcggctag ttgcttttgc gaggcctttg tctgaaggat gcaaaatcta
     3361 cggatgctag cgaggggggg aaggggggag agattacctc ataccatgtc gcttgcacca
     3421 atcaccactc tcctgtcgcg gcttctctgg gcagacggag gggtctggac caacaggaaa
     3481 aggccttggc ccatccccat ggtgaccgag ctgtatataa ggaggcgcat ccgcccaagt
     3541 gccgcaggtt ctcttacatc gaccgcttaa gagtcgcgct gtaagaagca acacctcctc
     3601 ctcgcctccg ccatccaccc ggcagccgcg aagcagcaac accggtgagc gggggatctc
     3661 gagctcaagc ttcgaattct gcagtcgact gccaccatgg ccgtcaatgt gtactccaca
     3721 tctgtgacca gtgaaaatct gagtcgccat gatatgcttg catgggtcaa cgactccctg
     3781 cacctcaact ataccaagat agaacagctt tgttcagggg cagcctactg ccagttcatg
     3841 gacatgctct tccccggctg tgtgcacttg aggaaagtga agttccaggc caaactagag
     3901 catgaataca tccacaactt caaggtgctg caagcagctt tcaagaagat gggtgttgac
     3961 aaaatcattc ctgtagagaa attagtgaaa ggaaaattcc aagataattt tgagtttatt
     4021 cagtggttta agaaattctt tgacgcaaac tatgatggaa aggattacaa ccctctgctg
     4081 gcgcggcagg gccaggacgt agcgccacct cctaacccag gtgatcagat cttcaacaaa
     4141 tccaagaaac tcattggcac agcagttcca cagaggacgt cccccacagg cccaaaaaac
     4201 atgcagacct ctggccggct gagcaatgtg gcccccccct gcattctccg gaagaatcct
     4261 ccatcagccc gaaatggcgg ccatgagact gatgcccaaa ttcttgaact caaccaacag
     4321 ctggtggact tgaagctgac agtggatggg ctggagaagg aacgtgactt ctacttcagc
     4381 aaacttcgtg acatcgagct catctgccag gagcatgaaa gtgaaaacag ccctgttatc
     4441 tcaggcatca ttggcatcct ctatgccaca gaggaaggat tcgcaccccc tgaggacgat
     4501 gagattgaag agcatcaaca agaagaccag gacgagtacc tggatcccgc caccatggtg
     4561 agcaagggcg aggaggataa catggcctct ctcccagcga cacatgagtt acacatcttt
     4621 ggctccatca acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa tccaaatgat
     4681 ggttatgagg agttaaacct gaagtccacc aagggtgacc tccagttctc cccctggatt
     4741 ctggtccctc atatcgggta tggcttccat cagtacctgc cctaccctga cgggatgtcg
     4801 cctttccagg ccgccatggt agatggctcc ggctaccaag tccatcgcac aatgcagttt
     4861 gaagatggtg cctcccttac tgttaactac cgctacacct acgagggaag ccacatcaaa
     4921 ggagaggccc aggtgaaggg gactggtttc cctgctgacg gtcctgtgat gaccaactcg
     4981 ctgaccgctg cggactggtg caggtcgaag aagacttacc ccaacgacaa aaccatcatc
     5041 agtaccttta agtggagtta caccactgga aatggcaagc gctaccggag cactgcgcgg
     5101 accacctaca cctttgccaa gccaatggcg gctaactatc tgaagaacca gccgatgtac
     5161 gtgttccgta agacggagct caagcactcc aagaccgagc tcaacttcaa ggagtggcaa
     5221 aaggccttta ccgatgtgat gggcatggac gagctgtaca agtaataggc ggccgcgact
     5281 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5341 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     5521 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5581 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5641 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5701 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5761 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5821 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5881 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5941 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     6001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6061 aata
//
