LOCUS       sequence-442624-        6400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual Cre/Dre-dependent expression of EB3-mNeon. Used as functional 
            (labelling of microtubule plus ends) Cre-Dre cotransfection 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  AUTHORS   Vinopal S, Dupraz S, Alfadil E, Pietralla T, Bendre S, Stiess M, 
            Falk S, Camargo Ortega G, Maghelli N, Tolic IM, Smejkal J, Gotz M, 
            Bradke F
  TITLE     Centrosomal microtubule nucleation regulates radial migration of 
            projection neurons independently of polarization in the developing 
            brain.
  JOURNAL   Neuron. 2023 Apr 19;111(8):1241-1263.e16. doi: 
            10.1016/j.neuron.2023.01.020. Epub 2023 Feb 15.
  PUBMED    36796357
REFERENCE   2  (bases 1 to 6400)
  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: 
            "10.1016/j.neuron.2023.01.020"; journalName: "Neuron"; date: 
            "2023-04-19- 19"; volume: "111"; issue: "8"; pages: "1241-1263.e16"
COMMENT     Sequence Label: pBactin-DIO-inv[roxSTOProx-EB3-mNeonGreen-bGH]
FEATURES             Location/Qualifiers
     source          1..6400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..126
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1927..2201
                     /label=chicken beta-actin promoter
     protein_bind    complement(3273..3306)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3357..3390)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3449..3673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3662..3679
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3695..4405)
                     /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      4141..4150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4402..4411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5260..5269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5316..5347
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    5376..5510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     gap             5757..5856
                     /estimated_length=100
     protein_bind    6026..6057
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    6103..6136
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    6187..6220
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6244..6260)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
       61 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
      121 gccagtgcca agttgggatc tttgcattgg cccacggctc tcaggatggg gatgctcccc
      181 ttcagcaccc ggttcccctt ggaaactgat ggtcctggct ctgtggcatg gcagtggcac
      241 tgtgaggagc cccctaccag cagcacacag tggggttggc actgccacgc tccggatgcc
      301 gcgctctgat ccaaccccat aatcaaggga acccgaattg ccccatcatt gcccccacca
      361 cccccatcct gccgggccct cacaccccac gctgccttct ggtgacattc cccagcccaa
      421 acccacggct tcatggctac cgcggggcat ttcccattgc cgccccatta tcagctctgc
      481 acacctcccg ctgtacccat gcctcgtggc tgcccttctt tgacgtataa tcttctaatt
      541 aatacccggc cttgtcaaag tggagcacaa acgttaatta attccccagc aggcaggtaa
      601 ttaacagtgt gactcccttt ttgctgcgag tggggctgat acagagagat gtggcactat
      661 ggagcccacg gggtcctggc actgggtgcc cacggaggtc cccatgtgct gcagtgtcac
      721 cgcctccgag gagacagtat tgtccctgcg gtgtccctgc agctcagctc tgtccacagg
      781 gccacctcca gtttggaggg gacacaatgc agccccgatg caacccatcc tcgcagcatc
      841 ccagggacaa agacctcact gcaagaccgc acacagggct gggtcccgct cccctaatat
      901 ctacagtgct tcttgcatgg ccccttaatc aatgcagtta atcagcatgc gctcatgcac
      961 cgctctggag ctgcaaagcc cctcgcagcg ctgctcacca acaccgcgca ccgccccggc
     1021 ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa tattgcccaa atgtaggcaa
     1081 aggcattcgg ctgccttgac ctccgccggg ccgggccctg cctgactcag ctccttactc
     1141 agcgctcgct tcctccctcc ggctgccacc gccgcagcgc acaccctgac aaagagtggc
     1201 ccttaacggg ctctgaggtg cacccagcag tgcactcagc agtccaaggg ccggcctgga
     1261 ggtttgcacc gctacgtgct gacattagca ttgaacttgg ccctgggtag tgctgcaggc
     1321 cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc cggtgcccct tcccctccct
     1381 tgcatcccag caggctgcac cccagcacca ggcccgtgca tgcatgctcc tggtgttatt
     1441 gcagcctggt gcatgcatgc gtcttgatgg tgcagcgctg tgcatgcatc ctccttggtg
     1501 tgtagcagtc tagtgcatgc atacccctcg gtgttattgc tgctctgtgc acgcacgctc
     1561 attgtatcac ttcatcccag tgcatgcact cacactggag cgattgctgc tcggtgcacg
     1621 cacactcatt gtatcactgc agctcagtgg ctgcacgcac accggtgtta ttgctgctcg
     1681 gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat gcacgctcat tgcccatcgc
     1741 tatccctgcc tctcctgctg gcgctccccg ggaggtgact tcaaggggac cgcaggacca
     1801 cctcgggggt ggggggaggg ctgcacacgc ggaccccgct ccccctcccc aacaaagcac
     1861 tgtggaatca aaaagggggg aggggggatg gaggggcgcg tcacaccccc gccccacacc
     1921 ctcacctcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     1981 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
     2041 gggggggcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
     2101 gtgcggcggc agccaatcag agcggcgcgc cccgaaagtt tccttttatg gcgaggcggc
     2161 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     2221 ttcgccccgt gccccgcccc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     2281 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     2341 tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa gggctccggg
     2401 agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
     2461 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     2521 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     2581 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     2641 gggggtgtgg gcgcggcggt cgggctgtaa cccccccctg cacccccctc cccgagttgc
     2701 tgagcacggc ccggcttcgg gtgcggggct ccgtgcgggg cgtggcgcgg ggctcgccgt
     2761 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     2821 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     2881 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     2941 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     3001 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     3061 gcgccgccgt ccccttctcc atctccagcc tcggggctgt ccgcaggggg acggctgcct
     3121 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gggtttatat
     3181 cttcccttct ctgttcctcc gcagccccca agctagcgtt cggcttctgg cgtgtgaccg
     3241 gcggctctag accaggcaga ctcgactccg gaataacttc gtataggata ctttatacga
     3301 agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa
     3361 cttcgtatag catacattat acgaagttat ggcgcgttac ttgtacagct agagcaacta
     3421 gctagcgagg ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg
     3481 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
     3541 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
     3601 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     3661 ctagaaggca cagtcgaggc tgatcagcga gctcttactt gtacagctcg tccatgccca
     3721 tcacatcggt aaaggccttt tgccactcct tgaagttgag ctcggtcttg gagtgcttga
     3781 gctccgtctt acggaacacg tacatcggct ggttcttcag atagttagcc gccattggct
     3841 tggcaaaggt gtaggtggtc cgcgcagtgc tccggtagcg cttgccattt ccagtggtgt
     3901 aactccactt aaaggtactg atgatggttt tgtcgttggg gtaagtcttc ttcgacctgc
     3961 accagtccgc agcggtcagc gagttggtca tcacaggacc gtcagcaggg aaaccagtcc
     4021 ccttcacctg ggcctctcct ttgatgtggc ttccctcgta ggtgtagcgg tagttaacag
     4081 taagggaggc accatcttca aactgcattg tgcgatggac ttggtagccg gagccatcta
     4141 ccatggcggc ctggaaaggc gacatcccgt cagggtaggg caggtactga tggaagccat
     4201 acccgatatg agggaccaga atccaggggg agaactggag gtcacccttg gtggacttca
     4261 ggtttaactc ctcataacca tcatttggat tgccggtgcc ctgacccacc atgtcaaagt
     4321 ccacaccgtt gatggagcca aagatgtgta actcatgtgt cgctgggaga gaggccatgt
     4381 tatcctcctc gcccttgctc accatggtgg cgggatccag gtactcgtcc tggtcttctt
     4441 gttgatgctc ttcaatctca tcgtcctcag ggggtgcgaa tccttcctct gtggcataga
     4501 ggatgccaat gatgcctgag ataacagggc tgttttcact ttcatgctcc tggcagatga
     4561 gctcgatgtc acgaagtttg ctgaagtaga agtcacgttc cttctccagc ccatccactg
     4621 tcagcttcaa gtccaccagc tgttggttga gttcaagaat ttgggcatca gtctcatggc
     4681 cgccatttcg ggctgatgga ggattcttcc ggagaatgca ggggggggcc acattgctca
     4741 gccggccaga ggtctgcatg ttttttgggc ctgtggggga cgtcctctgt ggaactgctg
     4801 tgccaatgag tttcttggat ttgttgaaga tctgatcacc tgggttagga ggtggcgcta
     4861 cgtcctggcc ctgccgcgcc agcagagggt tgtaatcctt tccatcatag tttgcgtcaa
     4921 agaatttctt aaaccactga ataaactcaa aattatcttg gaattttcct ttcactaatt
     4981 tctctacagg aatgattttg tcaacaccca tcttcttgaa agctgcttgc agcaccttga
     5041 agttgtggat gtattcatgc tctagtttgg cctggaactt cactttcctc aagtgcacac
     5101 agccggggaa gagcatgtcc atgaactggc agtaggctgc ccctgaacaa agctgttcta
     5161 tcttggtata gttgaggtgc agggagtcgt tgacccatgc aagcatatca tggcgactca
     5221 gattttcact ggtcacagat gtggagtaca cattgacggc catggtggca gtcgactgca
     5281 gaattcgatc tagcttgggc tgcagggcgg ctgcctaact ttaaataatg ccaattattt
     5341 aaagttaatt aagggttccg gatcagcttg atggggatcc agacatgata agatacattg
     5401 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5461 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5521 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     5581 aaaacctcta caaatgtggt atggctgatt atgatcctct agagtcgcag atccagacat
     5641 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5701 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaatannnn
     5761 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     5821 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnngtta acaacaacaa ttgcattcat
     5881 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     5941 aaatgtggta tggctgatta tgatcctcta gagtcgcaga tcctctagag tcgcagatct
     6001 gcaagctttc atttattcat cgcgataact ttaaataatg ccaattattt aaagttaggt
     6061 ggcgggtcga ccccctacgc gtcacgccta tggtggccta gcataacttc gtataaagta
     6121 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     6181 caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcggcc
     6241 gccactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     6301 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     6361 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
//
