LOCUS       sequence-391918-        7570 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 7570)
  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 7570)
  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: pCAG-DIO-inv[roxSTOProx-EB3 mNeonGreen-bGH]
FEATURES             Location/Qualifiers
     source          1..7570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    complement(1661..1694)
                     /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(1745..1778)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1837..2061
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2050..2067
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2083..2793)
                     /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      2529..2538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2790..2799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3648..3657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3704..3735
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    3764..3898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4018..4152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4272..4406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4573..4604
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    4650..4683
                     /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    4734..4767
                     /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(4791..4808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4791..4807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4800..4822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4949..5404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5036..5055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5246..5267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5430..5534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5535..5603,5604..6395)
                     /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(5753..5772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6566..7154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7055..7074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7308..7325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7442..7463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7478..7483,7484..7501,7502..7508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7516..7532
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7521..7543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7540..7556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7540..7556
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccrggs ggggsggggs ggggsgrggg
      541 gsggggsggg gsgrggsggr grggkgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1081 tgtgtgcgkg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1621 tgtgaccggc ggctctagac caggcagact cgactccgga ataacttcgt ataggatact
     1681 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     1741 cttaataact tcgtatagca tacattatac gaagttatgg cgcgttactt gtacagctag
     1801 agcaactagc tagcgaggtt ctttccgcct cagaagccat agagcccacc gcatccccag
     1861 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     1921 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     1981 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     2041 gctggcaact agaaggcaca gtcgaggctg atcagcgagc tcttacttgt acagctcgtc
     2101 catgcccatc acatcggtaa aggccttttg ccactccttg aagttgagct cggtcttgga
     2161 gtgcttgagc tccgtcttac ggaacacgta catcggctgg ttcttcagat agttagccgc
     2221 cattggcttg gcaaaggtgt aggtggtccg cgcagtgctc cggtagcgct tgccatttcc
     2281 agtggtgtaa ctccacttaa aggtactgat gatggttttg tcgttggggt aagtcttctt
     2341 cgacctgcac cagtccgcag cggtcagcga gttggtcatc acaggaccgt cagcagggaa
     2401 accagtcccc ttcacctggg cctctccttt gatgtggctt ccctcgtagg tgtagcggta
     2461 gttaacagta agggaggcac catcttcaaa ctgcattgtg cgatggactt ggtagccgga
     2521 gccatctacc atggcggcct ggaaaggcga catcccgtca gggtagggca ggtactgatg
     2581 gaagccatac ccgatatgag ggaccagaat ccagggggag aactggaggt cacccttggt
     2641 ggacttcagg tttaactcct cataaccatc atttggattg ccggtgccct gacccaccat
     2701 gtcaaagtcc acaccgttga tggagccaaa gatgtgtaac tcatgtgtcg ctgggagaga
     2761 ggccatgtta tcctcctcgc ccttgctcac catggtggcg ggatccaggt actcgtcctg
     2821 gtcttcttgt tgatgctctt caatctcatc gtcctcaggg ggtgcgaatc cttcctctgt
     2881 ggcatagagg atgccaatga tgcctgagat aacagggctg ttttcacttt catgctcctg
     2941 gcagatgagc tcgatgtcac gaagtttgct gaagtagaag tcacgttcct tctccagccc
     3001 atccactgtc agcttcaagt ccaccagctg ttggttgagt tcaagaattt gggcatcagt
     3061 ctcatggccg ccatttcggg ctgatggagg attcttccgg agaatgcagg ggggggccac
     3121 attgctcagc cggccagagg tctgcatgtt ttttgggcct gtgggggacg tcctctgtgg
     3181 aactgctgtg ccaatgagtt tcttggattt gttgaagatc tgatcacctg ggttaggagg
     3241 tggcgctacg tcctggccct gccgcgccag cagagggttg taatcctttc catcatagtt
     3301 tgcgtcaaag aatttcttaa accactgaat aaactcaaaa ttatcttgga attttccttt
     3361 cactaatttc tctacaggaa tgattttgtc aacacccatc ttcttgaaag ctgcttgcag
     3421 caccttgaag ttgtggatgt attcatgctc tagtttggcc tggaacttca ctttcctcaa
     3481 gtgcacacag ccggggaaga gcatgtccat gaactggcag taggctgccc ctgaacaaag
     3541 ctgttctatc ttggtatagt tgaggtgcag ggagtcgttg acccatgcaa gcatatcatg
     3601 gcgactcaga ttttcactgg tcacagatgt ggagtacaca ttgacggcca tggtggcagt
     3661 cgactgcaga attcgatcta gcttgggctg cagggcggct gcctaacttt aaataatgcc
     3721 aattatttaa agttaattaa gggttccgga tcagcttgat ggggatccag acatgataag
     3781 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3841 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3901 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3961 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag agtcgcagat
     4021 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4081 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4141 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4201 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct
     4261 ctagagtcgc agatccagac atgataagat acattgatga gtttggacaa accacaacta
     4321 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     4381 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4441 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     4501 ctgattatga tcctctagag tcgcagatcc tctagagtcg cagatctgca agctttcatt
     4561 tattcatcgc gataacttta aataatgcca attatttaaa gttaggtggc gggtcgaccc
     4621 cctacgcgtc acgcctatgg tggcctagca taacttcgta taaagtatcc tatacgaagt
     4681 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     4741 cgtataatgt atgctatacg aagttatgaa ttcgatatca agcggccgcc actggccgtc
     4801 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4861 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4921 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4981 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5041 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5101 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5161 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5221 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5281 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5341 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5401 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5461 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5521 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5581 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5641 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5701 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5761 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5821 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5881 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5941 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6001 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6061 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6121 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6181 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6241 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6301 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6361 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6421 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6481 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6541 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6601 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6661 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6721 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6781 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6841 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6901 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6961 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7021 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7081 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7141 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7201 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7261 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7321 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7381 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7441 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7501 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7561 aggcgcgccg
//
