LOCUS       sequence-390738-        7265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the centrosome targeting carboxy-terminal domain (CTD)
            of Cdk5rap2 fused to AcGFP. Used to displace endogenous Cdk5rap2 
            from centrosomes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7265)
  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 7265)
  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-AcGFP-Cdk5rap2-CTD
FEATURES             Location/Qualifiers
     source          1..7265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1179..1201
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1269..1985
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2661..2795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2698..2717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2752..2771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2809..2825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2809..2825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2822..2844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2833..2849
                     /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)."
     promoter        complement(join(2857..2863,2864..2881,2882..2887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2902..2923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3040..3057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3211..3799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3291..3310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3970..4761,4762..4830))
                     /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     4593..4612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4831..4935)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5003..5021
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5059..5081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5181..5200
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5394..5416
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5408..5425
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5409..5425
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(7226..7265,1..236)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
       61 gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgrg gggcggggcg
      121 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      181 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      241 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      301 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      361 gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc
      421 ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg
      481 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      541 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
      601 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
      661 gkgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
      721 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
      781 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
      841 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
      901 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
      961 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1021 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1081 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgtccgca
     1141 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1201 cggcggggtt tatatcttcc cttctctgtt cctccgcagc ccccaagcta gcgctaccgg
     1261 tcgccaccat ggtgagcaag ggcgccgagc tgttcaccgg catcgtgccc atcctgatcg
     1321 agctgaatgg cgatgtgaat ggccacaagt tcagcgtgag cggcgagggc gagggcgatg
     1381 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cctgtgccct
     1441 ggcccaccct ggtgaccacc ctgagctacg gcgtgcagtg cttctcacgc taccccgatc
     1501 acatgaagca gcacgacttc ttcaagagcg ccatgcctga gggctacatc caggagcgca
     1561 ccatcttctt cgaggatgac ggcaactaca agtcgcgcgc cgaggtgaag ttcgagggcg
     1621 ataccctggt gaatcgcatc gagctgaccg gcaccgattt caaggaggat ggcaacatcc
     1681 tgggcaataa gatggagtac aactacaacg cccacaatgt gtacatcatg accgacaagg
     1741 ccaagaatgg catcaaggtg aacttcaaga tccgccacaa catcgaggat ggcagcgtgc
     1801 agctggccga ccactaccag cagaataccc ccatcggcga tggccctgtg ctgctgcccg
     1861 ataaccacta cctgtccacc cagagcgccc tgtccaagga ccccaacgag aagcgcgatc
     1921 acatgatcta cttcggcttc gtgaccgccg ccgccatcac ccacggcatg gatgagctgt
     1981 acaagtccgg actcagatct cacatgctgt gcctgattga ggactatgat gccctctata
     2041 agcagatcag ctggggccag acactgcttg ccaagatgga tattcaaacc caagaggctc
     2101 tgagccccac aagtcagaag ctgggaccaa aggcttcatt ctctgtgcct ctgagcaagt
     2161 ttctctccag catgaacaca gccaagctga tcctggaaaa agcctccagg ttgctgaagc
     2221 tcttctggag ggtctctgtc cccaccaacg gccagtgttc ccttcactgt gaccagattg
     2281 gagaaatgaa ggcagagatc accaaactac acaaaaaatt gtttgaacaa gaaaagaagc
     2341 tgcagaacac agcaaaactt ctgcagcaga gcaagcacca ggagaaaatc atctttgatc
     2401 agttggtcat cacccaccaa gtccttcgga aagccagggg aaacctggag ctcaggccta
     2461 gagctgccca tccaggaaca tccagtccca gcagaccggg ctcatgagtc gacggtaccg
     2521 cgggcccggg atccaccgga tctagataag taatgatcat aatcagccat atcacatctg
     2581 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2641 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2701 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2761 ccaaactcat caatgtatct tatcatgtct ggatcaattc gtaatcatgt catagctgtt
     2821 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2881 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2941 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3001 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3061 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3121 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3181 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3241 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3541 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3601 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3661 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3721 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3781 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3841 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3961 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4141 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4201 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4261 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4321 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4381 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4441 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4501 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4561 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4621 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4681 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4741 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4801 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4861 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4921 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4981 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5041 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5101 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5161 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5221 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5281 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5341 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5401 acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc ccacggctct
     5461 caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg gtcctggctc
     5521 tgtggcatgg cagtggcact gtgaggagcc ccctaccagc agcacacagt ggggttggca
     5581 ctgccacgct ccggatgccg cgctctgatc caaccccata atcaagggaa cccgaattgc
     5641 cccatcattg cccccaccac ccccatcctg ccgggccctc acaccccacg ctgccttctg
     5701 gtgacattcc ccagcccaaa cccacggctt catggctacc gcggggcatt tcccattgcc
     5761 gccccattat cagctctgca cacctcccgc tgtacccatg cctcgtggct gcccttcttt
     5821 gacgtataat cttctaatta atacccggcc ttgtcaaagt ggagcacaaa cgttaattaa
     5881 ttccccagca ggcaggtaat taacagtgtg actccctttt tgctgcgagt ggggctgata
     5941 cagagagatg tggcactatg gagcccacgg ggtcctggca ctgggtgccc acggaggtcc
     6001 ccatgtgctg cagtgtcacc gcctccgagg agacagtatt gtccctgcgg tgtccctgca
     6061 gctcagctct gtccacaggg ccacctccag tttggagggg acacaatgca gccccgatgc
     6121 aacccatcct cgcagcatcc cagggacaaa gaccccactg caagaccgca cacagggctg
     6181 ggtcccgctc ccctaatatc tacagtgctt cttgcatggc cccttaatca atgcagttaa
     6241 tcagcatgcg ctcatgcacc gctctggagc tgcaaagccc ctcgcagcgc tgctcaccaa
     6301 caccgcgcac cgccccggcc cagcctgcag cacgcgctgc aaacaggaaa gaaacaaaat
     6361 attgcccaaa tgtaggcaaa ggcattcggc tgccttgacc tccgccgggc cgggccctgc
     6421 ctgactcagc tccttactca gcgctcgctt cctccctccg gctgccaccg ccgcagcgca
     6481 caccctgaca aagagtggcc cttaacgggc tctgaggtgc acccagcagt gcactcagca
     6541 gtccaagggc cggcctggag gtttgcaccg ctacgtgctg acattagcat tgaacttggc
     6601 cctgggtagt gctgcaggcc gggcggggtg ggtgtagaga gtgcagcgcg cgttgcaccc
     6661 ggtgcccctt cccctccctt gcatcccagc aggctgcacc ccagcaccag gcccgtgcat
     6721 gcatgctcct ggtgttattg cagcctggtg catgcatgcg tcttgatggt gcagcgctgt
     6781 gcatgcatcc tccttggtgt gtagcagtct agtgcatgca tacccctcgg tgttattgct
     6841 gctctgtgca cgcacgctca ttgtatcact tcatcccagt gcatgcactc acactggagc
     6901 gattgctgct cggtgcacgc acactcattg tatcactgca gctcagtggc tgcacgcaca
     6961 ccggtgttat tgctgctcgg tgcgtgcatg cacatcagtg tcgctgcagc tcagtgcatg
     7021 cacgctcatt gcccatcgct atccctgcct ctcctgctgg cgctccccgg gaggtgactt
     7081 caaggggacc gcaggaccac ctcgggggtg gggggagggc tgcacacgcg gaccccgctc
     7141 cccctcccca acaaagcact gtggaatcaa aaagggggga ggggggatgg aggggcgcgt
     7201 cacacccccg ccccacaccc tcacctcgag gtgagcccca cgttctgctt cactctcccc
     7261 atctc
//
