LOCUS       sequence-223600-        6509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human mutant DCX tagged with 2A peptide at it's C-terminus
            and mCherry in mammalian cells. Serine at position 306 substituted 
            for alanine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6509)
  AUTHORS   Slepak TI, Salay LD, Lemmon VP, Bixby JL
  TITLE     Dyrk kinases regulate phosphorylation of doublecortin, cytoskeletal 
            organization, and neuronal morphology.
  JOURNAL   Cytoskeleton (Hoboken). 2012 Jul;69(7):514-27. doi: 
            10.1002/cm.21021. Epub 2012 Mar 7.
  PUBMED    22359282
REFERENCE   2  (bases 1 to 6509)
  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: "10.1002/cm.21021"; 
            journalName: "Cytoskeleton (Hoboken)"; date: "2012-07"; volume: 
            "69"; issue: "7"; pages: "514-27"
COMMENT     Sequence Label: pAAV-DCX(S306A)-2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          876..1254
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             2453..2506
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2534..3241
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2676..2694)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2944..2963
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3256..3732
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3455..3474)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3772..3912
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3772..3901
                     /label=AAV2 ITR
     rep_origin      3987..4442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4074..4093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4284..4305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4459..4478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4578..4600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4638..4656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4724..4828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4829..4897,4898..5689)
                     /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(5047..5066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5860..6448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6349..6368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt ccccggggat ctaccatgga acttgatttt ggacactttg acgaaagaga
     1381 taagacatcc aggaacatgc gaggctcccg gatgaatggg ttgcctagcc ccactcacag
     1441 cgcccactgt agcttctacc gaaccagaac cttgcaggca ctgagtaatg agaagaaagc
     1501 caagaaggta cgtttctacc gcaatgggga ccgctacttc aaggggattg tgtacgctgt
     1561 gtcctctgac cgttttcgca gctttgacgc cttgctggct gacctgacgc gatctctgtc
     1621 tgacaacatc aacctgcctc agggagtgcg ttacatttac accattgatg gatccaggaa
     1681 gatcggaagc atggatgaac tggaggaagg ggaaagctat gtctgttcct cagacaactt
     1741 ctttaaaaag gtggagtaca ccaagaatgt caatcccaac tggtctgtca acgtaaaaac
     1801 atctgccaat atgaaagccc cccagtcctt ggctagcagc aacagtgcac aggccaggga
     1861 gaacaaggac tttgtgcgcc ccaagctggt taccatcatc cgcagtgggg tgaagcctcg
     1921 gaaggctgtg cgtgtgcttc tgaacaagaa gacagcccac tcttttgagc aagtcctcac
     1981 tgatatcaca gaagccatca aactggagac cggggttgtc aaaaaactct acactctgga
     2041 tggaaaacag gtaacttgtc tccatgattt ctttggtgat gatgatgtgt ttattgcctg
     2101 tggtcctgaa aaatttcgct atgctcagga tgatttttct ctggatgaaa atgaatgccg
     2161 agtcatgaag ggaaacccat cagccacagc tggcccaaag gcatccccaa cacctcagaa
     2221 gacttcagcc aagagccctg gtcctatgcg ccgaagcaag gctccagctg actcagcaaa
     2281 cggaacctcc agcagccagc tctctacccc caagtctaag cagtctccca tctctacgcc
     2341 caccagtcct ggcagcctcc ggaagcacaa ggacctgtac ctgcctctgt ccttggatga
     2401 ctcggactcg cttggtgatt ccatgtctag agtcgaccgg ggtaccaccg gtgaaggccg
     2461 cggcagcctg ctgacctgcg gcgatgtgga agaaaacccg ggcccggcgc cgggcagcac
     2521 tagtcggaag cttgtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     2581 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     2641 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     2701 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     2761 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     2821 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     2881 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     2941 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     3001 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     3061 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     3121 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     3181 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     3241 actcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3301 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3361 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3421 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3481 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     3541 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3601 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3661 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3721 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     3781 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3841 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3901 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3961 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4201 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4321 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4381 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4441 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4501 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4561 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4621 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4681 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5041 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5341 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5701 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6001 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6481 ggccttttgc tggccttttg ctcacatgt
//
