LOCUS       sequence-354019-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of mCherry and a shRNA against Ptbp1 under 
            the constitutive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  AUTHORS   Wang LL, Serrano C, Zhong X, Ma S, Zou Y, Zhang CL
  TITLE     Revisiting astrocyte to neuron conversion with lineage tracing in 
            vivo.
  JOURNAL   Cell. 2021 Oct 14;184(21):5465-5481.e16. doi: 
            10.1016/j.cell.2021.09.005. Epub 2021 Sep 27.
  PUBMED    34582787
REFERENCE   2  (bases 1 to 6557)
  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.cell.2021.09.005"; journalName: "Cell"; date: 
            "2021-10-14- 14"; volume: "184"; issue: "21"; pages: "5465-5481.e16"
COMMENT     Sequence Label: AAV-CAG-LSL-mCherry-shPtbp1
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..719
                     /label=chicken beta-actin promoter
     primer_bind     998..1020
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1082..1101
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1150..1183
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1207..1240
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1407..1541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1661..1795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1824..1857
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1889..1898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1895..2605
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2040..2058)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2308..2327
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           2803..2817
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    3001..3589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3054..3074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3472..3483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3632..3753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3915..4044
                     /label=AAV2 ITR
     rep_origin      complement(4281..4869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4361..4380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5040..5831,5832..5900))
                     /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     5663..5682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5901..6005)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6031..6486)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6168..6189)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6380..6399
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgat ttgataccgc
     1141 gggccctaag aagttcctat tctctagaaa gtataggaac ttcgtcgaca tttaaatcat
     1201 ttaaatataa cttcgtataa tgtatgctat acgaagttat tcgcgatgaa taaatgaaag
     1261 cttgcagatc tgcgactcta gaggatctgc gactctagag gatcataatc agccatacca
     1321 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1381 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1501 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgcgactct agaggatcat
     1561 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1621 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccat
     1801 caagctgatc cggaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     1861 gtccctcgac ctgcagccca agaccggtgc caccatggtg agcaagggcg aggaggataa
     1921 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1981 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2041 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     2101 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2161 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2221 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2281 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2341 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2401 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     2461 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     2521 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     2581 cggcatggac gagctgtaca agtagcggcc gcaagccttg ttaagtgctc gcttcggcag
     2641 cacatatact atgttgaatg aggcttcagt actttacaga atcgttgcct gcacatcttg
     2701 gaaacacttg ctgggattac ttcgacttct taacccaaca gaaggctcga gaaggtatat
     2761 tgctgttgac agtgagcgag ggtgaagatc ctgttcaata tagtgaagcc acagatgtat
     2821 attgaacagg atcttcaccc gtgcctactg cctcggactt caaggggcta gaattcgagc
     2881 aattatcttg tttactaaaa ctgaatacct tgctatctct ttgatacatt tttacaaagc
     2941 tgaattaaaa tggtataaat taaatcactt tggatccgtc gagggatcaa gcttatcgat
     3001 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3061 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3121 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3181 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3241 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3301 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3361 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3421 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3481 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3541 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     3601 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3661 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3721 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3781 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3841 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     3901 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3961 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4021 ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4081 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6001 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     6061 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6121 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6181 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6241 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6301 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6361 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6421 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6481 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6541 cttcgctatt acgccag
//
