LOCUS       sequence-409730-        5934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vDip-C AAV vector with mouse Pcp2 (also known as L7) promoter 
            driving green fluorophore mGreenLantern with a KASH nuclear membrane
            tag to isolate cell nuclei of cerebellar Purkinje cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  AUTHORS   Tan L, Shi J, Moghadami S, Parasar B, Wright CP, Seo Y, Vallejo K, 
            Cobos I, Duncan L, Chen R, Deisseroth K
  TITLE     Lifelong restructuring of 3D genome architecture in cerebellar 
            granule cells.
  JOURNAL   Science. 2023 Sep 8;381(6662):1112-1119. doi: 
            10.1126/science.adh3253. Epub 2023 Sep 7.
  PUBMED    37676945
REFERENCE   2  (bases 1 to 5934)
  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: 
            "10.1126/science.adh3253"; journalName: "Science"; date: "2023-09-8-
            8"; volume: "381"; issue: "6662"; pages: "1112-1119"
COMMENT     Sequence Label: vDip-C: pAAV-Pcp2-mGreenLantern-KASH
FEATURES             Location/Qualifiers
     source          1..5934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      995..1004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2061..2649
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2114..2134)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2532..2543)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2681..3157
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2880..2899)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3197..3337
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3197..3326
                     /label=AAV2 ITR
     rep_origin      3412..3867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3709..3730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3884..3903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4003..4025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4063..4081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4149..4253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4254..4322,4323..5114)
                     /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(4472..4491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5285..5873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5774..5793
                     /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 gcgtggttcc accctcatgt tggttgatct tcaacattgt
      181 actgacttct tcccacttga cattcctcaa ggtcctgtga tcatggctgg gtctagtgag
      241 gttcaaacct gcactgccct acccacaccc acacccagct cagcgtcagt caggatcaac
      301 aattacctag agatcatctt tctggggctt aagcattggt gggagcagat gggatatgag
      361 ctggggattt gggaatgggg gaagatatct gctccccctc cccctacacc ctagcctttt
      421 aaaaggcctt ctcaggtcag agaccaggag aaaagtatag gagagataca caatggacca
      481 ggaagaagaa aagggagagg gaggctcaga ccttctagac aaggtaagag ggctctggct
      541 gactccacca tccgcttctt gaggtctcgg cacctgtaat tgacaagatt aattcattta
      601 tagggcatct aattagcaag caagtctctg gagtcccctg acccagttac tataacacac
      661 agggggtata ggtaggagag tataagagcc cctcctcagg gcaaatgaat ggattcttag
      721 tactgtcccc caagagatag taggtactag gatttagggg cacttctgag ccccatttcc
      781 ctggtaagtg tcccaacccc ccaaatcaac ccaagcctgg tctcaatcta ggacagtggt
      841 agaatgctgt ccctagagtc agtaccatgt gaaattgtgc tgcaggcagg ggccccaggc
      901 tgggaggtgg gggttggggg agtcaggggc aggtcaggga aggagactca ggtttcattt
      961 agagaaattc tgcagacccg tgaggactgg atccgccacc atggttagta aaggcgaaga
     1021 gctgtttacc ggggtggtgc caattctagt ggaattggat ggcgatgtga atggacacaa
     1081 gtttagtgta cgcggcgaag gggaaggtga cgcgacaaat gggaaattga cactcaaatt
     1141 catttgcacc accgggaagc tcccagtgcc gtggccgacc cttgtgacga ccttggggta
     1201 tggtgttgcc tgctttgctc gctatccgga tcatatgaaa caacatgatt tcttcaaaag
     1261 tgctatgccc gaaggttatg tccaggagag aactatatca tttaaagatg atggaaccta
     1321 taaaacgagg gcagaggtca agtttgaggg agacactctg gtgaatcgga ttgtgctgaa
     1381 aggaatcgat ttcaaggaag atgggaatat actcggtcat aagttggagt acaacttcaa
     1441 ctcccacaag gtgtatataa ctgcggacaa acaaaagaat ggcataaaag caaattttaa
     1501 gacacggcat aacgtggaag acggcggcgt gcaactggcc gaccattacc agcaaaatac
     1561 accaatcggc gatggcccag tgctgttgcc ggataatcat taccttagtc accaatctaa
     1621 actatctaag gaccctaatg agaaacgcga tcacatggtg ttgaaagagc gcgtcacggc
     1681 agccgggatt acccatgaca tggacgaact gtataagagt ggcctgcgga gccgggaaga
     1741 agaagaagaa actgattcaa gaatgcctca tttggattca cctggatcaa gtcaaccaag
     1801 gcgatcattt ctttctcgcg taataagagc tgctctgcca ctgcaactct tgctcctgct
     1861 tttgctcctt ctcgcgtgtc ttctgcccgc tagtgaggac gattattcat gtactcaagc
     1921 aaataatttc gctcggtcat tttatcctat gctccgttat acgaatggtc ccccacctac
     1981 ttgatggcgc gccaatcaag gctaggtgga ggctcagtgg gccacctccc acctaagaat
     2041 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2101 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2161 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2221 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2281 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2341 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2401 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2461 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     2521 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2581 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2641 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     2701 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2761 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2821 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2881 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2941 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     3001 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3061 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3121 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     3181 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3241 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3301 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3361 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3421 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3481 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3541 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3601 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3661 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3721 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3781 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3841 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3901 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4021 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4081 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
