LOCUS       sequence-353989-        5829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry and a shRNA against Ptbp1 under the human GFAP
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5829)
  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 5829)
  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-hGFAP-mCherry-shPtbp1
FEATURES             Location/Qualifiers
     source          1..5829
                     /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      840..849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             846..1556
                     /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(991..1009)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1259..1278
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           1754..1768
                     /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    1952..2540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2005..2025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2423..2434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2576..3052
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3092..3232
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3092..3221
                     /label=AAV2 ITR
     rep_origin      3307..3762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3394..3413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3604..3625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3779..3798)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3898..3920
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3958..3976)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4044..4148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4149..4217,4218..5009)
                     /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"
     rep_origin      5180..5768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5669..5688
                     /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 gcgaattaat tcaacatatc ctggtgtgga gtaggggacg
      181 ctgctctgac agaggctcgg gggcctgagc tggctctgtg agctggggag gaggcagaca
      241 gccaggcctt gtctgcaagc agacctggca gcattgggct ggccgccccc cagggcctcc
      301 tcttcatgcc cagtgaatga ctcaccttgg cacagacaca atgttcgggg tgggcacagt
      361 gcctgcttcc cgccgcaccc cagcccccct caaatgcctt ccgagaagcc cattgagcag
      421 ggggcttgca ttgcacccca gcctgacagc ctggcatctt gggataaaag cagcacagcc
      481 ccctaggggc tgcccttgct gtgtggcgcc accggcggtg gagaacaagg ctctattcag
      541 cctgtgccca ggaaagggga tcaggggatg cccaggcatg gacagtgggt ggcagggggg
      601 gagaggaggg ctgtctgctt cccagaagtc caaggacaca aatgggtgag gggagagctc
      661 tccccatagc tgggctgcgg cccaacccca ccccctcagg ctatgccagg gggtgttgcc
      721 aggggcaccc gggcatcgcc agtctagccc actccttcat aaagccctcg catcccagga
      781 gcgagcagag ccagagcagg ttggagagga gacgcatcac ctccgctgct cgcaccggtg
      841 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      901 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      961 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1021 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     1081 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1141 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1201 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1261 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1321 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1381 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1441 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1501 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtagcggc
     1561 cgcaagcctt gttaagtgct cgcttcggca gcacatatac tatgttgaat gaggcttcag
     1621 tactttacag aatcgttgcc tgcacatctt ggaaacactt gctgggatta cttcgacttc
     1681 ttaacccaac agaaggctcg agaaggtata ttgctgttga cagtgagcga gggtgaagat
     1741 cctgttcaat atagtgaagc cacagatgta tattgaacag gatcttcacc cgtgcctact
     1801 gcctcggact tcaaggggct agaattcgag caattatctt gtttactaaa actgaatacc
     1861 ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa ttaaatcact
     1921 ttggatccgt cgagggatca agcttatcga taatcaacct ctggattaca aaatttgtga
     1981 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2041 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2101 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2161 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2221 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2281 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2341 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2401 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2461 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2521 cctttgggcc gcctccccgc atcgataccg agcgctgctc gatcgagaga tctacgggtg
     2581 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2641 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2701 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     2761 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     2821 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2881 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2941 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     3001 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3061 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3121 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3181 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3241 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3301 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3361 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3481 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3541 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3601 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3661 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3721 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3781 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3841 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3901 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3961 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4021 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4081 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4141 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4201 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4261 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4321 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4381 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4441 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4501 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4561 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4621 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4681 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4741 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4801 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4861 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4921 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4981 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5041 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5101 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5161 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5221 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5281 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5341 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5401 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5461 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5521 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5581 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5641 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5701 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5761 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5821 ctcacatgt
//
