LOCUS       sequence-391826-        7007 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of the gamma-tubulin-binding domain (gTBD) of NEDD1 fused
            to mOrange. 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 7007)
  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 7007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-mOrange2-NEDD1-gTBD
FEATURES             Location/Qualifiers
     source          1..7007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1179..1201
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1254..1263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1260..1967
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2403..2537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2440..2459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2494..2513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2551..2567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2564..2586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2575..2591
                     /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(2599..2605,2606..2623,2624..2629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2644..2665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2782..2799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2953..3541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3033..3052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3712..4503,4504..4572))
                     /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     4335..4354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4573..4677)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4745..4763
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4801..4823)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4923..4942
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5136..5158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5150..5167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5151..5167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6968..7007,1..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 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 cggcggggtt tatatcttcc cttctctgtt cctccgcagc ccccaagcta gcagccacca
     1261 tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1321 tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1381 gcccctacga gggctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct
     1441 tcgcctggga catcctgtcc cctcatttca cctacggctc caaggcctac gtgaagcacc
     1501 ccgccgacat ccccgactac ttcaagctgt ccttccccga gggcttcaag tgggagcgcg
     1561 tgatgaacta cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1621 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtga
     1681 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggtg
     1741 ccctgaaggg caagatcaag atgaggctga agctgaagga cggcggccac tacacctccg
     1801 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg
     1861 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1921 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtac tcagatctcg
     1981 agctcaagct tcgaattcta gccgacagca ttggaaataa ccggcaaaat gcaccattga
     2041 cttccattca aattcgtttt attcagaaca tgatacagga aacgttggat gactttagag
     2101 aagcatgcca tagggacatt gtgaatttgc aagtggagat gattaaacag tttcatatgc
     2161 aactgaatga aatgcattct ttgctggaaa gatactcagt gaatgaaggt ttagtggctg
     2221 aaattgaaag actacgagaa gaaaacaaaa gattacgggc ccacttttga ggatccaccg
     2281 gatctagata agtaatgatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     2341 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2401 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2461 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2521 cttatcatgt ctggatcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     2581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2701 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4681 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4741 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4801 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4861 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4921 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4981 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5041 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5101 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5161 ggccagtgcc aagttgggat ctttgcattg gcccacggct ctcaggatgg ggatgctccc
     5221 cttcagcacc cggttcccct tggaaactga tggtcctggc tctgtggcat ggcagtggca
     5281 ctgtgaggag ccccctacca gcagcacaca gtggggttgg cactgccacg ctccggatgc
     5341 cgcgctctga tccaacccca taatcaaggg aacccgaatt gccccatcat tgcccccacc
     5401 acccccatcc tgccgggccc tcacacccca cgctgccttc tggtgacatt ccccagccca
     5461 aacccacggc ttcatggcta ccgcggggca tttcccattg ccgccccatt atcagctctg
     5521 cacacctccc gctgtaccca tgcctcgtgg ctgcccttct ttgacgtata atcttctaat
     5581 taatacccgg ccttgtcaaa gtggagcaca aacgttaatt aattccccag caggcaggta
     5641 attaacagtg tgactccctt tttgctgcga gtggggctga tacagagaga tgtggcacta
     5701 tggagcccac ggggtcctgg cactgggtgc ccacggaggt ccccatgtgc tgcagtgtca
     5761 ccgcctccga ggagacagta ttgtccctgc ggtgtccctg cagctcagct ctgtccacag
     5821 ggccacctcc agtttggagg ggacacaatg cagccccgat gcaacccatc ctcgcagcat
     5881 cccagggaca aagaccccac tgcaagaccg cacacagggc tgggtcccgc tcccctaata
     5941 tctacagtgc ttcttgcatg gccccttaat caatgcagtt aatcagcatg cgctcatgca
     6001 ccgctctgga gctgcaaagc ccctcgcagc gctgctcacc aacaccgcgc accgccccgg
     6061 cccagcctgc agcacgcgct gcaaacagga aagaaacaaa atattgccca aatgtaggca
     6121 aaggcattcg gctgccttga cctccgccgg gccgggccct gcctgactca gctccttact
     6181 cagcgctcgc ttcctccctc cggctgccac cgccgcagcg cacaccctga caaagagtgg
     6241 cccttaacgg gctctgaggt gcacccagca gtgcactcag cagtccaagg gccggcctgg
     6301 aggtttgcac cgctacgtgc tgacattagc attgaacttg gccctgggta gtgctgcagg
     6361 ccgggcgggg tgggtgtaga gagtgcagcg cgcgttgcac ccggtgcccc ttcccctccc
     6421 ttgcatccca gcaggctgca ccccagcacc aggcccgtgc atgcatgctc ctggtgttat
     6481 tgcagcctgg tgcatgcatg cgtcttgatg gtgcagcgct gtgcatgcat cctccttggt
     6541 gtgtagcagt ctagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct
     6601 cattgtatca cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac
     6661 gcacactcat tgtatcactg cagctcagtg gctgcacgca caccggtgtt attgctgctc
     6721 ggtgcgtgca tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg
     6781 ctatccctgc ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc
     6841 acctcggggg tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca
     6901 ctgtggaatc aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac
     6961 cctcacctcg aggtgagccc cacgttctgc ttcactctcc ccatctc
//
