LOCUS       sequence-391833-        8777 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of neural precursor cell expressed developmentally 
            down-regulated 1 (Need1) fused to mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8777)
  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 8777)
  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-Nedd1-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..8777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1179..1201
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             3267..3977
                     /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      3522..3531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4173..4307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4210..4229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4264..4283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4321..4337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4321..4337)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4334..4356)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4345..4361
                     /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(4369..4375,4376..4393,4394..4399))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4414..4435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4552..4569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4723..5311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4803..4822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5482..6273,6274..6342))
                     /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     6105..6124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6343..6447)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6515..6533
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6571..6593)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6693..6712
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6906..6928
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6920..6937
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6921..6937
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(8738..8777,1..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 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgs
      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 tgcaggaaaa cctcagattt gcctcatcag gagatgatgt taaaatatgg gatgcttcgt
     1321 ttctgacatt ggtggataaa ttcaaccccc acacgtcacc gcatggaatc agttcaatct
     1381 gttggagcag caataataac ttcctagtga ctgcgtcttc tggtggtgac aaaatagtgg
     1441 tgtcaagttg caaatgtaag cccgttccac ttttggagct tgcagaaggg caaaagcaga
     1501 catgtgtcga tttaaattct acatcaatgt atttggcaag tggaggccta aataacactg
     1561 ttaatatttg ggatttaaag tcaaaacgac ttcatcggtc tcttaaggac cataaacggg
     1621 aagtaacttg tgtggcgtac aattggaatg attgttacat tgcttccggg tctctcagtg
     1681 gtgagattat cttacacagc gtaaccacta atacctccag tacgcctttc ggccatggga
     1741 gtaagcagcc tgtccggcac atcaagtact cgttgtttag gaagtctctc ttgggcagcg
     1801 tttcggataa tggagtagtg accctctggg atgtgaacag tcagagtgca taccacagct
     1861 ttgacagtac acacaaagcc ccagcttcag gcctctgctt ctctcctgtc aacgaactgc
     1921 tgtttgtgac cataggcctg gataaaagaa tcatcctgta tgacacttca agtaagaagc
     1981 tggtgaaaac tttagtggct gacactcctc tgactgctgt agattttatg ccagatggag
     2041 ccagtttggc tattggatcc tcccgtggga aaatatatca gtatgactta aggatgctga
     2101 aatctcccgt gaaaaccatc agcgcccaca agacatctgt gcagtgtata gcatttcagt
     2161 actccacatc tctgacgaag gcaagtttaa ataaaggctg ctcaaataag gccacagcag
     2221 cgaacaaacg gagtgttcct gtgggcagca gcagtggggc tgcccagaat tctggaattg
     2281 tcagagaagc acccagccct tccattgcca cagttctccc acagcccgtg acgacagctg
     2341 tgggcaaagg agcaggggct gctcaggacg aagcaggttt gcctcgaagc aaaaatacag
     2401 atatcttatc taaggaaact gatgctggaa aaagtcaaga cttctccagc tttgatgaca
     2461 cggggaagag tagtttaggg gacatgtttt caccaattag agatgatgct gtagtcaaca
     2521 aaggaggtga cgagtctgta ggcaaaggag atggccttga ctttctacca cagctaaact
     2581 cggtgttccc cctgaggaag aatgcaggct cctccagcac tctggtgctg cactccagcc
     2641 cttttaatgt ccttatgggg acagggaagg aagagcatga gagtcacgag ctgtcagccg
     2701 agtctaagaa ggcacacctt gggaagccgg agcctaagga ctccttgaag cagtttacaa
     2761 agttgatctc ctctggtgct gaacctggaa ttctaaacac ctgtccatca tctaaccaag
     2821 caagaaatct tgaaaagttt gaaaagccag aaaaagatat tgaagtccag ttgatgcacg
     2881 aaccctcagt caatggaccc tcaaccccag taccaaaggc agcctcctcc gtcactgctg
     2941 gagtagccag ttcactctca gaaaaaatag tcgacaccct tgggaacagt cggccggccg
     3001 cgccacttac ctctgtccag atccgcttca ttcagaacat gatacaagaa accctggatg
     3061 acttcaggga agcttgccat agggacattg tgaatctgca agtggaaatg attaaacagt
     3121 ttcatatgca actgaatgaa atgcactctt tgctggaaag atactcagtg aatgaaggct
     3181 tagtggctga gatcgaaagg ctgcgagaag aaaacaaaag gctgcgggcc cacttctcaa
     3241 gcctcctcga cggaagtggt ggatccatgg tgagcaaggg cgaggaggat aacatggcct
     3301 ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg gactttgaca
     3361 tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac ctgaagtcca
     3421 ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg tatggcttcc
     3481 atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg gtagatggct
     3541 ccggctacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt actgttaact
     3601 accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgaag gggactggtt
     3661 tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg tgcaggtcga
     3721 agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt tacaccactg
     3781 gaaatggcaa gcgctaccgg agcactgcgc ggaccaccta cacctttgcc aagccaatgg
     3841 cggctaacta tctgaagaac cagccgatgt acgtgttccg taagacggag ctcaagcact
     3901 ccaagaccga gctcaacttc aaggagtggc aaaaggcctt taccgatgtg atgggcatgg
     3961 acgagctgta caagtaagcg gccgccatat gcatcctagg cctattaata ttccggagta
     4021 tacgtagccg gctaacgtta acaaccggta cctctagata agtaatgatc ataatcagcc
     4081 atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4141 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4201 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4261 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat tcgtaatcat
     4321 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4381 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4441 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4501 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4561 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4621 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4681 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4741 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4801 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4861 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4921 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4981 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5041 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5101 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5161 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5221 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5281 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5341 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5401 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5461 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5521 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5581 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5641 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5701 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5761 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5821 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5881 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5941 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6001 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6061 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6121 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6181 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6241 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6301 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6361 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6421 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6481 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6541 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6601 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6661 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6721 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6781 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6841 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6901 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagttgggat ctttgcattg
     6961 gcccacggct ctcaggatgg ggatgctccc cttcagcacc cggttcccct tggaaactga
     7021 tggtcctggc tctgtggcat ggcagtggca ctgtgaggag ccccctacca gcagcacaca
     7081 gtggggttgg cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg
     7141 aacccgaatt gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca
     7201 cgctgccttc tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca
     7261 tttcccattg ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg
     7321 ctgcccttct ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca
     7381 aacgttaatt aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga
     7441 gtggggctga tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc
     7501 ccacggaggt ccccatgtgc tgcagtgtca ccgcctccga ggagacagta ttgtccctgc
     7561 ggtgtccctg cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg
     7621 cagccccgat gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg
     7681 cacacagggc tgggtcccgc tcccctaata tctacagtgc ttcttgcatg gccccttaat
     7741 caatgcagtt aatcagcatg cgctcatgca ccgctctgga gctgcaaagc ccctcgcagc
     7801 gctgctcacc aacaccgcgc accgccccgg cccagcctgc agcacgcgct gcaaacagga
     7861 aagaaacaaa atattgccca aatgtaggca aaggcattcg gctgccttga cctccgccgg
     7921 gccgggccct gcctgactca gctccttact cagcgctcgc ttcctccctc cggctgccac
     7981 cgccgcagcg cacaccctga caaagagtgg cccttaacgg gctctgaggt gcacccagca
     8041 gtgcactcag cagtccaagg gccggcctgg aggtttgcac cgctacgtgc tgacattagc
     8101 attgaacttg gccctgggta gtgctgcagg ccgggcgggg tgggtgtaga gagtgcagcg
     8161 cgcgttgcac ccggtgcccc ttcccctccc ttgcatccca gcaggctgca ccccagcacc
     8221 aggcccgtgc atgcatgctc ctggtgttat tgcagcctgg tgcatgcatg cgtcttgatg
     8281 gtgcagcgct gtgcatgcat cctccttggt gtgtagcagt ctagtgcatg catacccctc
     8341 ggtgttattg ctgctctgtg cacgcacgct cattgtatca cttcatccca gtgcatgcac
     8401 tcacactgga gcgattgctg ctcggtgcac gcacactcat tgtatcactg cagctcagtg
     8461 gctgcacgca caccggtgtt attgctgctc ggtgcgtgca tgcacatcag tgtcgctgca
     8521 gctcagtgca tgcacgctca ttgcccatcg ctatccctgc ctctcctgct ggcgctcccc
     8581 gggaggtgac ttcaagggga ccgcaggacc acctcggggg tggggggagg gctgcacacg
     8641 cggaccccgc tccccctccc caacaaagca ctgtggaatc aaaaaggggg gaggggggat
     8701 ggaggggcgc gtcacacccc cgccccacac cctcacctcg aggtgagccc cacgttctgc
     8761 ttcactctcc ccatctc
//
