LOCUS       sequence-391838-        7787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Pericentrin-AKAP450 centrosomal targeting (PACT) 
            domain fused to mOrange2 and human (h) Centrosomin motif 1 (CM1). 
            PACT targets CM1 to the centrosome .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7787)
  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 7787)
  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-PACT-mOrange2-hCM1
FEATURES             Location/Qualifiers
     source          1..7787
                     /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             2103..2810
                     /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    3183..3317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3220..3239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3274..3293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3331..3347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3344..3366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3355..3371
                     /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(3379..3385,3386..3403,3404..3409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3424..3445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3562..3579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4492..5283,5284..5352))
                     /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     5115..5134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5353..5457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5525..5543
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5581..5603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5703..5722
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5916..5938
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5930..5947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5931..5947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(7748..7787,1..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 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 gcagccacca
     1261 tggctgacat cgaagccata atcgcttcag aaaaggaagt ctggaacaga gaaaagctat
     1321 ctcttcagaa agcgttgaaa agggcagagg cgaaagtgta taaactgaaa gctgaactgc
     1381 gcaatgatgc tttgctgcgg agtctgagcc ctgactctga gcatgccgcc ctgcagaaaa
     1441 tttataacaa atacctgaga gcttcaagtt tccgcaaggc tctcatatat cagaaaaaat
     1501 acctcctgct gctcctgggg ggcttccagg agtgtgagga cgccaccctg ggggttctgg
     1561 cccgaatggg gggacaccca gccttgaaag atcccaagac aatcaccagc cacccgaagg
     1621 ggttcacccg gttccgctct gccgtcagag tgtctattgc catctcaaga atgaaatttt
     1681 tggtgcggcg gtggcagcaa gtgacaagta ctagctccat caatattaac agggatggct
     1741 ttggcctgag tccaggcatt gaaaagactg acccattcta tcattcgtct ggtgggctgg
     1801 agctgtatgg agaaccaaga cacacaacat accgctcaag atttgatctg gactacccta
     1861 ggtctccctt gccattgcag aataggtacc cgggtgctcc agacttaaat tctacttcaa
     1921 tggcttcttc tcaacttcac cagtacaatc ctgataaatc tctgacagac tatgtcactc
     1981 ggctagaggc actgaaaagg cgacttggag ctatacagtc agggtcgact actcaatttc
     2041 attttggcat gaggagaaag cttggtggag gcagcgcatc aggcggaagc ggaagcagat
     2101 ctatggtgag caagggcgag gagaataaca tggccatcat caaggagttc atgcgcttca
     2161 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2221 gccgccccta cgagggcttt cagaccgcta agctgaaggt gaccaagggt ggccccctgc
     2281 ccttcgcctg ggacatcctg tcccctcatt tcacctacgg ctccaaggcc tacgtgaagc
     2341 accccgccga catccccgac tacttcaagc tgtccttccc cgagggcttc aagtgggagc
     2401 gcgtgatgaa ctacgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2461 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2521 tgatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2581 gtgccctgaa gggcaagatc aagatgaggc tgaagctgaa ggacggcggc cactacacct
     2641 ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacatcg
     2701 tcgacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2761 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tctggcacag
     2821 gtggaagcat cgatacagtg tctcccacca gagcacggaa catgaaggac tttgaaaatc
     2881 aaatcactga attgaagaaa gaaaacttta acctaaagct ccgcatctat ttccttgagg
     2941 aaagaatgca acaggaattt catggcccca ctgaacatat ctactaagcg gccgccatat
     3001 gcatcctagg cctattaata ttccggagta tacgtagccg gctaacgtta acaaccggta
     3061 cctctagata agtaatgatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     3121 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3181 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3241 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3301 cttatcatgt ctggatcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     3361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5461 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5521 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5581 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5641 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5701 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5761 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5821 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5881 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5941 ggccagtgcc aagttgggat ctttgcattg gcccacggct ctcaggatgg ggatgctccc
     6001 cttcagcacc cggttcccct tggaaactga tggtcctggc tctgtggcat ggcagtggca
     6061 ctgtgaggag ccccctacca gcagcacaca gtggggttgg cactgccacg ctccggatgc
     6121 cgcgctctga tccaacccca taatcaaggg aacccgaatt gccccatcat tgcccccacc
     6181 acccccatcc tgccgggccc tcacacccca cgctgccttc tggtgacatt ccccagccca
     6241 aacccacggc ttcatggcta ccgcggggca tttcccattg ccgccccatt atcagctctg
     6301 cacacctccc gctgtaccca tgcctcgtgg ctgcccttct ttgacgtata atcttctaat
     6361 taatacccgg ccttgtcaaa gtggagcaca aacgttaatt aattccccag caggcaggta
     6421 attaacagtg tgactccctt tttgctgcga gtggggctga tacagagaga tgtggcacta
     6481 tggagcccac ggggtcctgg cactgggtgc ccacggaggt ccccatgtgc tgcagtgtca
     6541 ccgcctccga ggagacagta ttgtccctgc ggtgtccctg cagctcagct ctgtccacag
     6601 ggccacctcc agtttggagg ggacacaatg cagccccgat gcaacccatc ctcgcagcat
     6661 cccagggaca aagaccccac tgcaagaccg cacacagggc tgggtcccgc tcccctaata
     6721 tctacagtgc ttcttgcatg gccccttaat caatgcagtt aatcagcatg cgctcatgca
     6781 ccgctctgga gctgcaaagc ccctcgcagc gctgctcacc aacaccgcgc accgccccgg
     6841 cccagcctgc agcacgcgct gcaaacagga aagaaacaaa atattgccca aatgtaggca
     6901 aaggcattcg gctgccttga cctccgccgg gccgggccct gcctgactca gctccttact
     6961 cagcgctcgc ttcctccctc cggctgccac cgccgcagcg cacaccctga caaagagtgg
     7021 cccttaacgg gctctgaggt gcacccagca gtgcactcag cagtccaagg gccggcctgg
     7081 aggtttgcac cgctacgtgc tgacattagc attgaacttg gccctgggta gtgctgcagg
     7141 ccgggcgggg tgggtgtaga gagtgcagcg cgcgttgcac ccggtgcccc ttcccctccc
     7201 ttgcatccca gcaggctgca ccccagcacc aggcccgtgc atgcatgctc ctggtgttat
     7261 tgcagcctgg tgcatgcatg cgtcttgatg gtgcagcgct gtgcatgcat cctccttggt
     7321 gtgtagcagt ctagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct
     7381 cattgtatca cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac
     7441 gcacactcat tgtatcactg cagctcagtg gctgcacgca caccggtgtt attgctgctc
     7501 ggtgcgtgca tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg
     7561 ctatccctgc ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc
     7621 acctcggggg tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca
     7681 ctgtggaatc aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac
     7741 cctcacctcg aggtgagccc cacgttctgc ttcactctcc ccatctc
//
