LOCUS       sequence-391815-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the centrosome targeting carboxy-terminal domain (CTD)
            laking the centrosome binding domain (deltaCBD) of Cdk5rap2 fused to
            AcGFP. It cannot displace Cdk5rap2 from centrosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  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 7235)
  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-CTDdeltaCBD
FEATURES             Location/Qualifiers
     source          1..7235
                     /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    2631..2765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2668..2687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2722..2741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2779..2795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2779..2795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2792..2814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2803..2819
                     /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(2827..2833,2834..2851,2852..2857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2872..2893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3010..3027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3181..3769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3261..3280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3940..4731,4732..4800))
                     /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     4563..4582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4801..4905)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4973..4991
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5029..5051)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5151..5170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5364..5386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5378..5395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5379..5395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(7196..7235,1..236)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
       61 gggggcgggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag 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 gggctgcgrg 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 cggcggggkt 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 agttgagggg aaacctggag ctcaggccta gagctgccca tccaggaaca tccagtccca
     2461 gcagaccggg ctcatgagtc gacggtaccg cgggcccggg atccaccgga tctagataag
     2521 taatgatcat aatcagccat atcacatctg tagaggtttt acttgcttta aaaaacctcc
     2581 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2641 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2701 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2761 ggatcaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3601 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3841 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3901 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3961 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4021 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4081 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4141 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4201 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4261 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4321 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4381 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4441 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4501 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4561 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4621 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4681 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4741 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4801 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4861 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4921 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4981 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5041 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5101 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5161 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5221 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5281 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5341 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     5401 gttgggatct ttgcattggc ccacggctct caggatgggg atgctcccct tcagcacccg
     5461 gttccccttg gaaactgatg gtcctggctc tgtggcatgg cagtggcact gtgaggagcc
     5521 ccctaccagc agcacacagt ggggttggca ctgccacgct ccggatgccg cgctctgatc
     5581 caaccccata atcaagggaa cccgaattgc cccatcattg cccccaccac ccccatcctg
     5641 ccgggccctc acaccccacg ctgccttctg gtgacattcc ccagcccaaa cccacggctt
     5701 catggctacc gcggggcatt tcccattgcc gccccattat cagctctgca cacctcccgc
     5761 tgtacccatg cctcgtggct gcccttcttt gacgtataat cttctaatta atacccggcc
     5821 ttgtcaaagt ggagcacaaa cgttaattaa ttccccagca ggcaggtaat taacagtgtg
     5881 actccctttt tgctgcgagt ggggctgata cagagagatg tggcactatg gagcccacgg
     5941 ggtcctggca ctgggtgccc acggaggtcc ccatgtgctg cagtgtcacc gcctccgagg
     6001 agacagtatt gtccctgcgg tgtccctgca gctcagctct gtccacaggg ccacctccag
     6061 tttggagggg acacaatgca gccccgatgc aacccatcct cgcagcatcc cagggacaaa
     6121 gaccccactg caagaccgca cacagggctg ggtcccgctc ccctaatatc tacagtgctt
     6181 cttgcatggc cccttaatca atgcagttaa tcagcatgcg ctcatgcacc gctctggagc
     6241 tgcaaagccc ctcgcagcgc tgctcaccaa caccgcgcac cgccccggcc cagcctgcag
     6301 cacgcgctgc aaacaggaaa gaaacaaaat attgcccaaa tgtaggcaaa ggcattcggc
     6361 tgccttgacc tccgccgggc cgggccctgc ctgactcagc tccttactca gcgctcgctt
     6421 cctccctccg gctgccaccg ccgcagcgca caccctgaca aagagtggcc cttaacgggc
     6481 tctgaggtgc acccagcagt gcactcagca gtccaagggc cggcctggag gtttgcaccg
     6541 ctacgtgctg acattagcat tgaacttggc cctgggtagt gctgcaggcc gggcggggtg
     6601 ggtgtagaga gtgcagcgcg cgttgcaccc ggtgcccctt cccctccctt gcatcccagc
     6661 aggctgcacc ccagcaccag gcccgtgcat gcatgctcct ggtgttattg cagcctggtg
     6721 catgcatgcg tcttgatggt gcagcgctgt gcatgcatcc tccttggtgt gtagcagtct
     6781 agtgcatgca tacccctcgg tgttattgct gctctgtgca cgcacgctca ttgtatcact
     6841 tcatcccagt gcatgcactc acactggagc gattgctgct cggtgcacgc acactcattg
     6901 tatcactgca gctcagtggc tgcacgcaca ccggtgttat tgctgctcgg tgcgtgcatg
     6961 cacatcagtg tcgctgcagc tcagtgcatg cacgctcatt gcccatcgct atccctgcct
     7021 ctcctgctgg cgctccccgg gaggtgactt caaggggacc gcaggaccac ctcgggggtg
     7081 gggggagggc tgcacacgcg gaccccgctc cccctcccca acaaagcact gtggaatcaa
     7141 aaagggggga ggggggatgg aggggcgcgt cacacccccg ccccacaccc tcacctcgag
     7201 gtgagcccca cgttctgctt cactctcccc atctc
//
