LOCUS       sequence-353987-        6588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NEUROD1-K139D and mCherry under the human GFAP 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  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 6588)
  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-ND1-K139D-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1920..1973
                     /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"
     regulatory      1980..1989
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1986..2696
                     /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(2131..2149)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2399..2418
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2715..3303
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2768..2788)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3186..3197)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3335..3811
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3534..3553)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3851..3991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3851..3980
                     /label=AAV2 ITR
     rep_origin      4066..4521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4153..4172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4363..4384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4538..4557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4657..4679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4717..4735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4803..4907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4908..4976,4977..5768)
                     /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      5939..6527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6428..6447
                     /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 gcgaattcaa catatcctgg tgtggagtag gggacgctgc
      181 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      241 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      301 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      361 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      421 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      481 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      541 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      601 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      661 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
      721 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
      781 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca ccgggatccg
      841 ccaccatgac caaatcgtac agcgagagtg ggctgatggg cgagcctcag ccccaaggtc
      901 ctccaagctg gacagacgag tgtctcagtt ctcaggacga ggagcacgag gcagacaaga
      961 aggaggacga cctcgaagcc atgaacgcag aggaggactc actgaggaac gggggagagg
     1021 aggaggacga agatgaggac ctggaagagg aggaagaaga ggaagaggag gatgacgatc
     1081 aaaagcccaa gagacgcggc cccaaaaaga agaagatgac taaggctcgc ctggagcgtt
     1141 ttaaattgag acgcatgaag gctaacgccc gggagcggaa ccgcatgcac ggactgaacg
     1201 cggcgctaga caacctgcgc aaggtggtgc cttgctattc taagacgcag aagctgtccg
     1261 atatcgagac tctgcgcttg gccaagaact acatctgggc tctgtcggag atcctgcgct
     1321 caggcaaaag cccagacctg gtctccttcg ttcagacgct ttgcaagggc ttatcccaac
     1381 ccaccaccaa cctggttgcg ggctgcctgc aactcaatcc tcggactttt ctgcctgagc
     1441 agaaccagga catgcccccc cacctgccga cggccagcgc ttccttccct gtacacccct
     1501 actcctacca gtcgcctggg ctgcccagtc cgccttacgg taccatggac agctcccatg
     1561 tcttccacgt taagcctccg ccgcacgcct acagcgcagc gctggagccc ttctttgaaa
     1621 gccctctgac tgattgcacc agcccttcct ttgatggacc cctcagcccg ccgctcagca
     1681 tcaatggcaa cttctctttc aaacacgaac cgtccgccga gtttgagaaa aattatgcct
     1741 ttaccatgca ctatcctgca gcgacactgg caggggccca aagccacgga tcaatcttct
     1801 caggcaccgc tgcccctcgc tgcgagatcc ccatagacaa tattatgtcc ttcgatagcc
     1861 attcacatca tgagcgagtc atgagtgccc agctcaatgc catatttcat gatgctagcg
     1921 aggggagagg aagtcttctg acctgcggag acgtcgaaga gaatcctgga cccaccggtg
     1981 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     2041 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     2101 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     2161 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     2221 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     2281 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2341 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2401 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2461 atggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2521 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2581 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2641 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtagctcg
     2701 agaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2761 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2821 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2881 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2941 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc ggaactttcg
     3001 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3061 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3121 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3181 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3241 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3301 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3361 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3421 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3481 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3541 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3601 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3661 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     3721 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3781 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3841 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3901 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3961 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4021 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4141 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4201 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4261 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4381 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4441 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4501 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4561 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4621 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4681 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4741 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4801 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4861 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5101 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5281 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5461 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5521 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5761 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5821 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5881 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5941 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6001 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6061 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6121 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6181 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6241 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6301 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6361 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6421 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6481 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6541 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
