LOCUS       sequence-353948-        6588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NEUROD1 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-NEUROD1-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   34..163
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1953..2006
                     /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      2013..2022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2019..2729
                     /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(2164..2182)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2432..2451
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2748..3336
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2801..2821)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3219..3230)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3368..3844
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3567..3586)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3884..4024
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3884..4013
                     /label=AAV2 ITR
     rep_origin      4099..4554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4186..4205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4396..4417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4571..4590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4690..4712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4750..4768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4836..4940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4941..5009,5010..5801)
                     /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      5972..6560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6461..6480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
      121 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgaatt
      181 caacatatcc tggtgtggag taggggacgc tgctctgaca gaggctcggg ggcctgagct
      241 ggctctgtga gctggggagg aggcagacag ccaggccttg tctgcaagca gacctggcag
      301 cattgggctg gccgcccccc agggcctcct cttcatgccc agtgaatgac tcaccttggc
      361 acagacacaa tgttcggggt gggcacagtg cctgcttccc gccgcacccc agcccccctc
      421 aaatgccttc cgagaagccc attgagcagg gggcttgcat tgcaccccag cctgacagcc
      481 tggcatcttg ggataaaagc agcacagccc cctaggggct gcccttgctg tgtggcgcca
      541 ccggcggtgg agaacaaggc tctattcagc ctgtgcccag gaaaggggat caggggatgc
      601 ccaggcatgg acagtgggtg gcaggggggg agaggagggc tgtctgcttc ccagaagtcc
      661 aaggacacaa atgggtgagg ggagagctct ccccatagct gggctgcggc ccaaccccac
      721 cccctcaggc tatgccaggg ggtgttgcca ggggcacccg ggcatcgcca gtctagccca
      781 ctccttcata aagccctcgc atcccaggag cgagcagagc cagagcaggt tggagaggag
      841 acgcatcacc tccgctgctc gcaccgggat ccgccaccat gaccaaatcg tacagcgaga
      901 gtgggctgat gggcgagcct cagccccaag gtcctccaag ctggacagac gagtgtctca
      961 gttctcagga cgaggagcac gaggcagaca agaaggagga cgacctcgaa gccatgaacg
     1021 cagaggagga ctcactgagg aacgggggag aggaggagga cgaagatgag gacctggaag
     1081 aggaggaaga agaggaagag gaggatgacg atcaaaagcc caagagacgc ggccccaaaa
     1141 agaagaagat gactaaggct cgcctggagc gttttaaatt gagacgcatg aaggctaacg
     1201 cccgggagcg gaaccgcatg cacggactga acgcggcgct agacaacctg cgcaaggtgg
     1261 tgccttgcta ttctaagacg cagaagctgt ccaaaatcga gactctgcgc ttggccaaga
     1321 actacatctg ggctctgtcg gagatcctgc gctcaggcaa aagcccagac ctggtctcct
     1381 tcgttcagac gctttgcaag ggcttatccc aacccaccac caacctggtt gcgggctgcc
     1441 tgcaactcaa tcctcggact tttctgcctg agcagaacca ggacatgccc ccccacctgc
     1501 cgacggccag cgcttccttc cctgtacacc cctactccta ccagtcgcct gggctgccca
     1561 gtccgcctta cggtaccatg gacagctccc atgtcttcca cgttaagcct ccgccgcacg
     1621 cctacagcgc agcgctggag cccttctttg aaagccctct gactgattgc accagccctt
     1681 cctttgatgg acccctcagc ccgccgctca gcatcaatgg caacttctct ttcaaacacg
     1741 aaccgtccgc cgagtttgag aaaaattatg cctttaccat gcactatcct gcagcgacac
     1801 tggcaggggc ccaaagccac ggatcaatct tctcaggcac cgctgcccct cgctgcgaga
     1861 tccccataga caatattatg tccttcgata gccattcaca tcatgagcga gtcatgagtg
     1921 cccagctcaa tgccatattt catgatgcta gcgaggggag aggaagtctt ctgacctgcg
     1981 gagacgtcga agagaatcct ggacccaccg gtgccaccat ggtgagcaag ggcgaggagg
     2041 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     2101 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2161 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     2221 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2281 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2341 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2401 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     2461 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2521 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2581 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2641 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2701 ccggcggcat ggacgagctg tacaagtagc tcgagaagct tatcgataat caacctctgg
     2761 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2821 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2881 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2941 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3001 ccaccacctg tcagctcctt tccggaactt tcgctttccc cctccctatt gccacggcgg
     3061 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3121 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3181 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3241 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3301 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     3361 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3421 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3481 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3541 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     3601 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     3661 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     3721 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     3781 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     3841 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3901 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3961 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4021 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4441 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4621 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4681 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4741 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4801 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5101 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5521 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5761 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggt
//
