LOCUS       sequence-391816-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the gamma-tubulin-binding domain (gTBD) of NEDD1 fused
            to enhanced (E) GFP. Used to displace endogenous gamma-TuRC from the
            centrosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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-EGFP-NEDD1-gTBD
FEATURES             Location/Qualifiers
     source          1..7025
                     /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             join(1269..1271,1272..1274,1275..1985)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1269..1271,1272..1274,1275..1985)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1314..1335)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1575..1594)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1922..1943
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2421..2555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2458..2477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2512..2531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2569..2585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2582..2604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2593..2609
                     /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(2617..2623,2624..2641,2642..2647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2662..2683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2800..2817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3051..3070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3730..4521,4522..4590))
                     /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     4353..4372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4591..4695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4763..4781
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4819..4841)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4941..4960
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5154..5176
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5168..5185
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5169..5185
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6986..7025,1..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 gtgggggggt 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 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1321 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1381 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1441 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1501 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1561 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1621 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1681 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1741 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1801 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1861 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1921 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1981 acaagtactc agatctcgag ctcaagcttc gaattctagc cgacagcatt ggaaataacc
     2041 ggcaaaatgc accattgact tccattcaaa ttcgttttat tcagaacatg atacaggaaa
     2101 cgttggatga ctttagagaa gcatgccata gggacattgt gaatttgcaa gtggagatga
     2161 ttaaacagtt tcatatgcaa ctgaatgaaa tgcattcttt gctggaaaga tactcagtga
     2221 atgaaggttt agtggctgaa attgaaagac tacgagaaga aaacaaaaga ttacgggccc
     2281 acttttgagg atccaccgga tctagataag taatgatcat aatcagccat atcacatctg
     2341 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2401 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2521 ccaaactcat caatgtatct tatcatgtct ggatcaattc gtaatcatgt catagctgtt
     2581 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2641 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2761 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3661 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3841 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4321 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4681 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4741 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4801 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4861 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4921 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     4981 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5041 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5101 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5161 acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc ccacggctct
     5221 caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg gtcctggctc
     5281 tgtggcatgg cagtggcact gtgaggagcc ccctaccagc agcacacagt ggggttggca
     5341 ctgccacgct ccggatgccg cgctctgatc caaccccata atcaagggaa cccgaattgc
     5401 cccatcattg cccccaccac ccccatcctg ccgggccctc acaccccacg ctgccttctg
     5461 gtgacattcc ccagcccaaa cccacggctt catggctacc gcggggcatt tcccattgcc
     5521 gccccattat cagctctgca cacctcccgc tgtacccatg cctcgtggct gcccttcttt
     5581 gacgtataat cttctaatta atacccggcc ttgtcaaagt ggagcacaaa cgttaattaa
     5641 ttccccagca ggcaggtaat taacagtgtg actccctttt tgctgcgagt ggggctgata
     5701 cagagagatg tggcactatg gagcccacgg ggtcctggca ctgggtgccc acggaggtcc
     5761 ccatgtgctg cagtgtcacc gcctccgagg agacagtatt gtccctgcgg tgtccctgca
     5821 gctcagctct gtccacaggg ccacctccag tttggagggg acacaatgca gccccgatgc
     5881 aacccatcct cgcagcatcc cagggacaaa gaccccactg caagaccgca cacagggctg
     5941 ggtcccgctc ccctaatatc tacagtgctt cttgcatggc cccttaatca atgcagttaa
     6001 tcagcatgcg ctcatgcacc gctctggagc tgcaaagccc ctcgcagcgc tgctcaccaa
     6061 caccgcgcac cgccccggcc cagcctgcag cacgcgctgc aaacaggaaa gaaacaaaat
     6121 attgcccaaa tgtaggcaaa ggcattcggc tgccttgacc tccgccgggc cgggccctgc
     6181 ctgactcagc tccttactca gcgctcgctt cctccctccg gctgccaccg ccgcagcgca
     6241 caccctgaca aagagtggcc cttaacgggc tctgaggtgc acccagcagt gcactcagca
     6301 gtccaagggc cggcctggag gtttgcaccg ctacgtgctg acattagcat tgaacttggc
     6361 cctgggtagt gctgcaggcc gggcggggtg ggtgtagaga gtgcagcgcg cgttgcaccc
     6421 ggtgcccctt cccctccctt gcatcccagc aggctgcacc ccagcaccag gcccgtgcat
     6481 gcatgctcct ggtgttattg cagcctggtg catgcatgcg tcttgatggt gcagcgctgt
     6541 gcatgcatcc tccttggtgt gtagcagtct agtgcatgca tacccctcgg tgttattgct
     6601 gctctgtgca cgcacgctca ttgtatcact tcatcccagt gcatgcactc acactggagc
     6661 gattgctgct cggtgcacgc acactcattg tatcactgca gctcagtggc tgcacgcaca
     6721 ccggtgttat tgctgctcgg tgcgtgcatg cacatcagtg tcgctgcagc tcagtgcatg
     6781 cacgctcatt gcccatcgct atccctgcct ctcctgctgg cgctccccgg gaggtgactt
     6841 caaggggacc gcaggaccac ctcgggggtg gggggagggc tgcacacgcg gaccccgctc
     6901 cccctcccca acaaagcact gtggaatcaa aaagggggga ggggggatgg aggggcgcgt
     6961 cacacccccg ccccacaccc tcacctcgag gtgagcccca cgttctgctt cactctcccc
     7021 atctc
//
