LOCUS       sequence-190876-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome that expresses nuclear localized mRuby2 from the mouse
            Dlx5/6 enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  AUTHORS   Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, 
            Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
  TITLE     Engineered AAVs for efficient noninvasive gene delivery to the 
            central and peripheral nervous systems.
  JOURNAL   Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 
            2017 Jun 26.
  PUBMED    28671695
REFERENCE   2  (bases 1 to 5519)
  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: "10.1038/nn.4593"; 
            journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue: 
            "8"; pages: "1172-1179"
COMMENT     Sequence Label: pAAV-mDlx-NLS-mRuby2
FEATURES             Location/Qualifiers
     source          1..5519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          849..981
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     964..983
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1078..1098
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1108..1818
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    1865..2453
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1918..1938)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2336..2347)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2622..2703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2659..2678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2713..2732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2782..2922
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2782..2911
                     /label=AAV2 ITR
     rep_origin      2997..3452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3084..3103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3294..3315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3469..3488)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3588..3610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3648..3666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3734..3838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3839..3907,3908..4699)
                     /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(4057..4076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4870..5458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5359..5378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagaattcta tacactcaca gtggtttggc
      181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt
      241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc
      301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc
      361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
      421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
      481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
      541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
      601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
      661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
      781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
      841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta
     1021 gtagccacca tgtctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     1081 aaaaagaaga gaaaggtaga agaccccatg gtgtctaagg gcgaagagct gatcaaggaa
     1141 aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca
     1201 ggtgaaggag aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag
     1261 ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt
     1321 acttttatca agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt
     1381 tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac
     1441 accagccttg aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc
     1501 tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg
     1561 tatccagcag atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt
     1621 ggccatctgt cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc
     1681 aagatgcccg gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat
     1741 gaaatgttcg tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg
     1801 atggacgagc tgtacaagta aggcgcgcca cccctgcagg gaattcgata tcaagcttat
     1861 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2041 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2101 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2161 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2221 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2281 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2341 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2401 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     2461 ccgagcgctg ctcgagagat ctgtgatagc ggccatcaag ctggccgcga ctctagatca
     2521 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2581 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2641 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2701 tgcattctag ttgtggtttg tccaaactca tcaatgtatc agcttatcga taccgcatgc
     2761 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2821 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2881 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2941 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3061 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3181 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3301 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3361 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3421 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3481 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3661 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3841 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3901 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3961 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4021 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4081 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4141 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4201 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4261 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4321 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4381 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4441 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4501 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4561 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4621 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4681 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
