LOCUS       sequence-392227-        8838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron-specific expression of human (h)Centrin fused to mNeonGreen. 
            Used in combination with Talpha1-Dre-pA plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8838)
  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 8838)
  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-rox-inv[Talpha1-iCre-pA]-rox-mNeonGreen-hCentrin1
FEATURES             Location/Qualifiers
     source          1..8838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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    1729..1760
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     2098..2117
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2116..2171
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2217..2236
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(3269..3289)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3292..3301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4410..4441
                     /label=rox
                     /bound_moiety="Dre recombinase"
     regulatory      4458..4467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4464..5171
                     /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      4719..4728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5757..5774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5763..5987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6059..6076)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6059..6075)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6068..6090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6217..6672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6304..6323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6514..6535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6698..6802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6803..6871,6872..7663)
                     /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(7021..7040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7834..8422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8323..8342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8576..8593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8710..8731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8746..8751,8752..8769,8770..8776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8784..8800
                     /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     8789..8811
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8808..8824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8808..8824
                     /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 gsgrggsgga 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 ccgccaccta actttaaata
     1741 attggcatta tttaaagtta tcgcgatgaa taaatgaaag ctgtcgaggg atcttcataa
     1801 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     1861 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatataaa
     1921 aaaaatctaa cctcaagtca aggcttttct atggaataag gaatggacag cagggggctg
     1981 tttcatatac tgatgacctc tttatagcca acctttgttc atggcagcca gcatatgggc
     2041 atatgttgcc aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc
     2101 atatgtcctt ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa
     2161 tttcctttat tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt
     2221 ggcagaggga aaaagatctt cagtccccat cctcgagcag cctcaccatg gccccagtct
     2281 cagagtccag gtttctgatg tagttcatca caatgttcac attggtccag ccaccagcct
     2341 gcatgatttc agggatggac acaccagccc tggccatgtc cctggcagca cccactctgg
     2401 cagagtggcc agaccaggcc aggtatctct gcccagagtc atccttggca ccatagatca
     2461 ggcggtgggt ggcctcaaag atcccttcca gggcccgggt ggacagttgg gaggtggcag
     2521 aaggggcagc cacaccattc tttctgaccc ggcagaacag gtagttgttg gggtcatcag
     2581 ccacaccaga cacagagatc catctctcca ccagcttggt aacccccagg gacagggcct
     2641 tctccacacc agctgtggac accagggtct tggtcctgcc aatgtggatc agcattctcc
     2701 caccatcggt gcgggagatg tccttcactc tgattctggc aatttcggca atgcgcagca
     2761 gggtgttgta ggcaatgccc aggaaggcca ggttcctgat gtcctggcat ctgtcagagt
     2821 tctccatcag ggatctgact tggtcaaagt cagtgcgttc aaaggccagg gcctgcttgg
     2881 ctctctcccc agcatccaca ttctcctttc tgattctcct catcaccagg gacacagcat
     2941 tggagtcaga agggcgaggc aggccagatc tcctgtgcag catgttgagc tggcccaggt
     3001 gctgttggat ggtcttcaca gccaggcctc tggcttgcag gtacaggagg tagtccctca
     3061 catcctcagg ttcagcaggg aaccatttcc tgttgttcag cttgcaccag gcagcccagg
     3121 atctgcacac agacaggagc atcttccagg tgtgttcaga gaaggcctgc ctgtccctga
     3181 acatgtccat caggttcttc ctgacttcat cagaggtggc atccacaggg agggcaggca
     3241 ggttttggtg cacagtcagc aggttggaga ctttcctctt cttcttgggc accatggtgg
     3301 cgaattcgtt gctgcttcgc ggctgccggg tggatggcgg aggcgaggag gaggtgttgc
     3361 ttcttacagc gcgactctta agcggtcgat gtaagagaac ctgcggcact tgggcggatg
     3421 cgcctcctta tatacagctc ggtcaccatg gggatgggcc aaggcctttt cctgttggtc
     3481 cagacccctc cgtctgccca gagaagccgc gacaggagag tggtgattgg tgcaagcgac
     3541 atggtatgag gtaatctctc cccccttccc cccctcgcta gcatccgtag attttgcatc
     3601 cttcagacaa aggcctcgca aaagcaacta gccgagcatc cattatgggg tggagctggg
     3661 gaggtgtaga gagggaagaa agaaaacggg ggttgggtga ttatacggag cttaaacctt
     3721 aaactgtagt taggataatt taaggttttc cccaacactt aatcacagaa tttgtgttga
     3781 gattctttat ggtaaattcc agcttttcat ttatactaca ctaaatttaa atacgttccg
     3841 tcaggattca acatctctgt tcgctttaaa taaagtagca agagctatgg gaatgatggt
     3901 caaaaaggcg tgggtaaatg tttagtttta cagttgtatt attgtggtta ttattattac
     3961 tattattatt aatatttgag caaaattcag aagctcagta tgaatagacc acgcctatga
     4021 ccctcaacag cacccgggag gcagaggcag gaagacttgc agctcaaggt catcataacc
     4081 gcccttagtg gtttatgcct gtaaatccca gcattgggaa gcggaggact gcggttattt
     4141 tgtacccggc ctggatatat agtgtgatat cctgtctcaa aaaccaaaaa caaacaaaga
     4201 gagagagaga gagaaaggaa aaaagagaaa caatatacaa agtagtactt aacgctttac
     4261 tagagagctt gctttcttta gcggtagtgc tctcccatca cccctccccg ctcgccccct
     4321 tacccgaccc tacctcaccc cagagcctgt gttattttgt ccctagaaat gagccatccc
     4381 tcctaatacg gaattccgga acccttaatt aactttaaat aattggcatt atttaaagtt
     4441 aggcagccgc cctgcaggcc accatggtga gcaagggcga ggaggataac atggcctctc
     4501 tcccagcgac acatgagtta cacatctttg gctccatcaa cggtgtggac tttgacatgg
     4561 tgggtcaggg caccggcaat ccaaatgatg gttatgagga gttaaacctg aagtccacca
     4621 agggtgacct ccagttctcc ccctggattc tggtccctca tatcgggtat ggcttccatc
     4681 agtacctgcc ctaccctgac gggatgtcgc ctttccaggc cgccatggta gatggctccg
     4741 gctaccaagt ccatcgcaca atgcagtttg aagatggtgc ctcccttact gttaactacc
     4801 gctacaccta cgagggaagc cacatcaaag gagaggccca ggtgaagggg actggtttcc
     4861 ctgctgacgg tcctgtgatg accaactcgc tgaccgctgc ggactggtgc aggtcgaaga
     4921 agacttaccc caacgacaaa accatcatca gtacctttaa gtggagttac accactggaa
     4981 atggcaagcg ctaccggagc actgcgcgga ccacctacac ctttgccaag ccaatggcgg
     5041 ctaactatct gaagaaccag ccgatgtacg tgttccgtaa gacggagctc aagcactcca
     5101 agaccgagct caacttcaag gagtggcaaa aggcctttac cgatgtgatg ggcatggacg
     5161 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     5221 ccatggcttc cggcttcaag aagcccagcg ctgcctccac cggccaaaag agaaaggtgg
     5281 cacctaagcc cgagctcact gaggatcaga agcaagaagt tcgggaagca tttgacctct
     5341 tcgacgtgga cggaagtggg accatcgacg cgaaggagct gaaggtggcc atgagagcgc
     5401 tgggcttcga acccaggaag gaagagatga agaaaatgat ctccgaggtg gacagggaag
     5461 gcacggggaa gatcagcttc aatgacttcc tggccgtgat gacgcagaag atgtccgaga
     5521 aggacaccaa agaagaaatc ctgaaggcct tcaggctctt tgatgacgat gagaccggga
     5581 agatctcgtt caaaaacctg aagcgtgtgg ccaacgagct gggggagaac ctcacggatg
     5641 aggagctgca ggagatgatc gacgaagctg atcgggatgg ggacggcgaa gtgaacgagg
     5701 aggagttcct tcggatcatg aagaagacca gcctttactg agagctcgct gatcagcctc
     5761 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5821 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5881 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5941 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     6001 aagaaccagc tggggctcga gatccactag ttctagcctc gaggctagag cggccgccac
     6061 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6121 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6181 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     6241 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6301 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6361 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6421 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6481 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6541 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6601 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6661 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6721 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6781 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6841 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6901 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6961 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7021 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7081 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7141 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7201 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7261 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7321 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7381 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7441 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7501 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7561 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7621 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7741 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7981 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8401 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8461 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8521 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8581 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8641 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8701 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8761 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8821 tgaccatgag gcgcgccg
//
