LOCUS       sequence-392232-        8597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron-specific expression of fragment 128-425 from A-Kinase 
            Anchoring Protein 9 (Akap9) fused to 2xHA-tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  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 8597)
  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-128-425...
FEATURES             Location/Qualifiers
     source          1..8597
                     /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(5516..5533)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5522..5746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5818..5835)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5818..5834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5827..5849)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5976..6431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6063..6082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6273..6294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6457..6561
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6562..6630,6631..7422)
                     /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(6780..6799)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7593..8181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8082..8101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8335..8352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8469..8490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8505..8510,8511..8528,8529..8535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8543..8559
                     /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     8548..8570
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8567..8583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8567..8583
                     /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 gsgrggcgga gaggkgcggc 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 gtacctccgg aggtggtgga ggtatggggg aaaacgagtt
     4621 ggctgagaag cagcatgagg tcgaagagtt gactcaggag ttggaagaaa tgagggccag
     4681 ctttgggacg gaagggctga agcagttaca agaattcgaa gccgccatta aacaaagaga
     4741 tggcatcatc actcagctca ctgctaactt gcagcaagcc agaaaagaaa aggatgacac
     4801 aacgatagaa tttttagagt tgacagaaca gagtcaaaag ctgcagattc agttccagca
     4861 cttacaggct aatgaaaccc tgcaaaacag cactcttagt cgaactgcaa cagatttact
     4921 gcaggctaaa cgacagatct ttactctgca gcaacagctg caagattatc agaaaaagga
     4981 ggaggacgtc caagcacaaa tttctttttt acaagagaaa attagaatgt tggaaatgga
     5041 aaaagacaga aaagtggaaa gcttaaatac aaaagaaata gaggaaaagc aggcaatcat
     5101 tgaggagttg aatacaagag taatagaaga agagagaaaa actgctgagc tagagaataa
     5161 agtgacaact gctgatgaat tactaggaca gctccaagga caggttacag aaaaggatca
     5221 ggagatcaaa agcctgaaac tagaactggt caactctcag cagaacgagc ggaagtgttc
     5281 agaggaaatc aaagagctaa tgaggacagt tgaagaactc cagaagagaa atctcaagga
     5341 cagccagctg gggaccggca ccgtgcgaag ggtggaacaa gaaacacaac ggaaactaag
     5401 ccttctccag gcagagctgg atgagatgta tgggaagcag atagtgcaaa tgaaacaaga
     5461 attaatgaaa cagcacatgt ctcagataga ggagctttga gagctcgctg atcagcctcg
     5521 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     5761 agaaccagct ggggctcgag atccactagt tctagcctcg aggctagagc ggccgccact
     5821 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5881 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5941 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     6001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6181 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6301 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6421 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6481 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6541 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6601 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6661 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6721 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6781 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6841 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6901 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6961 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7021 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7081 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7141 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7201 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7261 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7321 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7381 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8281 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8341 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8401 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8461 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8521 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8581 gaccatgagg cgcgccg
//
