LOCUS       sequence-392064-        8201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron-specific expression of human (h)Centrin fused to 2xHA-tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  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 8201)
  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: pCAG-lox-inv[Talpha1-iCre-pA]-lox-2xHA-hCentrin1
FEATURES             Location/Qualifiers
     source          1..8201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..664
                     /label=chicken beta-actin promoter
     intron          665..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1605..1627
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1694..1712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1741..1774
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2112..2131
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2130..2185
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2231..2250
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(3283..3303)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3306..3315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4441..4474
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4502..4528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4532..4558
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5120..5137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5126..5350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5422..5439)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5422..5438)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5431..5453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5579..6034)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5716..5737)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5928..5947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6061..6165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6166..6234,6235..7026)
                     /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     complement(6384..6403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7197..7785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7686..7705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7939..7956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8073..8094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8109..8114,8115..8132,8133..8139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8147..8163
                     /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)."
     primer_bind     8152..8174
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8171..8187
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8171..8187
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccrggs ggggsggggs ggggsgrggg
      541 gsggggsggg gsgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
      961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1081 tgtgtgcgkg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1621 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gatccttaat taataatacg actcactata gggggtcgac ccgccaccaa tcatttaaat
     1741 ataacttcgt ataatgtatg ctatacgaag ttattcgcga tgaataaatg aaagctgtcg
     1801 agggatcttc ataagagaag agggacagct atgactggga gtagtcagga gaggaggaaa
     1861 aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa aaataacaca
     1921 aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg
     1981 acagcagggg gctgtttcat atactgatga cctctttata gccaaccttt gttcatggca
     2041 gccagcatat gggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa
     2101 tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt
     2161 gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg
     2221 tccccataat ttttggcaga gggaaaaaga tcttcagtcc ccatcctcga gcagcctcac
     2281 catggcccca gtctcagagt ccaggtttct gatgtagttc atcacaatgt tcacattggt
     2341 ccagccacca gcctgcatga tttcagggat ggacacacca gccctggcca tgtccctggc
     2401 agcacccact ctggcagagt ggccagacca ggccaggtat ctctgcccag agtcatcctt
     2461 ggcaccatag atcaggcggt gggtggcctc aaagatccct tccagggccc gggtggacag
     2521 ttgggaggtg gcagaagggg aagccacacc attctttctg acccggcaga acaggtagtt
     2581 gttggggtca tcagccacac cagacacaga gatccatctc tccaccagct tggtaacccc
     2641 cagggacagg gccttctcca caccagctgt ggacaccagg gtcttggtcc tgccaatgtg
     2701 gatcagcatt ctcccaccat cggtgcggga gatgtccttc actctgattc tggcaatttc
     2761 ggcaatgcgc agcagggtgt tgtaggcaat gcccaggaag gccaggttcc tgatgtcctg
     2821 gcatctgtca gagttctcca tcagggatct gacttggtca aagtcagtgc gttcaaaggc
     2881 cagggcctgc ttggctctct ccccagcatc cacattctcc tttctgattc tcctcatcac
     2941 cagggacaca gcattggagt cagaagggcg aggcaggcca gatctcctgt gcagcatgtt
     3001 gagctggccc aggtgctgtt ggatggtctt cacagccagg cctctggctt gcaggtacag
     3061 gaggtagtcc ctcacatcct caggttcagc agggaaccat ttcctgttgt tcagcttgca
     3121 ccaggcagcc caggatctgc acacagacag gagcatcttc caggtgtgtt cagagaaggc
     3181 ctgcctgtcc ctgaacatgt ccatcaggtt cttcctgact tcatcagagg tggcatccac
     3241 agggagggca ggcaggtttt ggtgcacagt cagcaggttg gagactttcc tcttcttctt
     3301 gggcaccatg gtggcgaatt cgttgctgct tcgcggctgc cgggtggatg gcggaggcga
     3361 ggaggaggtg ttgcttctta cagcgcgact cttaagcggt cgatgtaaga gaacctgcgg
     3421 cacttgggcg gatgcgcctc cttatataca gctcggtcac catggggatg ggccaaggcc
     3481 ttttcctgtt ggtccagacc cctccgtctg cccagagaag ccgcgacagg agagtggtga
     3541 ttggtgcaag cgacatggta tgaggtaatc tctcccccct tccccccctc gctagcatcc
     3601 gtagattttg catccttcag acaaaggcct cgcaaaagca actagccgag catccattat
     3661 ggggtggagc tggggagggg tagagaggga agaaagaaaa cgggggttgg gtgattatac
     3721 ggagcttaaa ccttaaactg tagttaggat aatttaaggt tttccccaac acttaatcac
     3781 agaatttgtg ttgagattct ttatggtaaa ttccagcttt tcatttatac tacactaaat
     3841 ttaaatacgt tccgtcagga ttcaacatct ctgttcgctt taaataaagt agcaagagct
     3901 atgggaatga tggtcaaaaa ggcgtgggta aatgtttagt tttacagttg tattattgtg
     3961 gttattatta ttactattat tattaatatt tgagcaaaat tcagaagctc agtatgaata
     4021 gaccacgcct atgaccctca acagcacccg ggaggcagag gcaggaagac ttgcagctca
     4081 aggtcatcat aaccgccctt agtggtttat gcctgtaaat cccagcattg ggaagcggag
     4141 gactgcggtt attttgtacc cggcctggat atatagtgtg atatcctgtc tcaaaaacca
     4201 aaaacaaaca aagagagaga gagagagaaa ggaaaaaaga gaaacaatat acaaagtagt
     4261 acttaacgct ttactagaga gcttgctttc tttagcggta gtgctctccc atcacccctc
     4321 cccgctcgcc cccttacccg accctacctc accccagagc ctgtgttatt ttgtccctag
     4381 aaatgagcca tccctcctaa tacggaatga tccccatcaa gctgatccgg aacccttaat
     4441 ataacttcgt ataatgtatg ctatacgaag ttattaggtc cctcgacctg cagccaccat
     4501 gtacccatac gatgttccag attacgctgg ctatccctat gacgtcccgg actatgcagg
     4561 aagttcagga ggttctagtg gtaccatggc ttccggcttc aagaagccca gcgctgcctc
     4621 caccggccaa aagagaaagg tggcacctaa gcccgagctc actgaggatc agaagcaaga
     4681 agttcgggaa gcatttgacc tcttcgacgt ggacggaagt gggaccatcg acgcgaagga
     4741 gctgaaggtg gccatgagag cgctgggctt cgaacccagg aaggaagaga tgaagaaaat
     4801 gatctccgag gtggacaggg aaggcacggg gaagatcagc ttcaatgact tcctggccgt
     4861 gatgacgcag aagatgtccg agaaggacac caaagaagaa atcctgaagg ccttcaggct
     4921 ctttgatgac gatgagaccg ggaagatctc gttcaaaaac ctgaagcgtg tggccaacga
     4981 gctgggggag aacctcacgg atgaggagct gcaggagatg atcgacgaag ctgatcggga
     5041 tggggacggc gaagtgaacg aggaggagtt ccttcggatc atgaagaaga ccagccttta
     5101 ctgagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5161 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5221 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5281 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5341 ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagatccac tagttctagc
     5401 ctcgaggcta gagcggccgc cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5461 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5521 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa
     5581 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5641 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5701 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5761 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5821 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5881 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5941 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6001 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     6061 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6181 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6241 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6301 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6361 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6421 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6481 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6541 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6601 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6661 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6721 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6781 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6841 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6901 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6961 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7021 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7081 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7321 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7981 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8041 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8101 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8161 caatttcaca caggaaacag ctatgaccat gaggcgcgcc g
//
