LOCUS       sequence-370990-        5829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of mCherry under the constitutive 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-CAG-FLEX-mCherry
FEATURES             Location/Qualifiers
     source          1..5829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     polyA_signal    292..413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(329..348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     383..402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    456..1044
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             562..573
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     971..991
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1086..1119
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1190..1223
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             1255..1308
                     /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      1315..1324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1321..2031
                     /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(1466..1484)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1734..1753
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2052..2085)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2148..2181)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2216..2235)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2297..2319)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2598..2876)
                     /label=chicken beta-actin promoter
     repeat_region   3187..3316
                     /label=AAV2 ITR
     rep_origin      3388..3843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3475..3494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3685..3706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3869..3973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3974..4042,4043..4834)
                     /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(4192..4211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5005..5593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5494..5513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
      241 ccacatctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
      301 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      361 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      421 tgctcgaagc ggccgcccgg gtcgacggta tcgatgcggg gaggcggccc aaagggagat
      481 ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc
      541 gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg
      601 cagaatccag gtggcaacac aggcgagcag ccaaggaaag gacgatgatt tccccgacaa
      661 caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
      721 ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg
      781 acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac
      841 gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata
      901 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
      961 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     1021 aattttgtaa tccagaggtt gattatcgat aagcttgata tcgaattcga acctcttcga
     1081 gggacataac ttcgtataat gtatgctata cgaagttatg ggtcgatggt gagatctgga
     1141 ctagagggtc gatggtgatg cttggatcaa aataggaaga atcccaatga taacttcgta
     1201 taaagtatcc tatacgaagt tatactagaa ttcggtaccg agctcgcagc tagcgagggg
     1261 agaggaagtc ttctgacctg cggagacgtc gaagagaatc ctggacccac cggtgccacc
     1321 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1381 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1441 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1501 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1561 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     1621 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1681 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1741 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1801 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1861 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1921 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1981 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta gctcgagggg
     2041 ggatccctag tataacttcg tatagcatac attatacgaa gttatgggtc gatggtgatg
     2101 cttggcaatt cgggtcgatg gtgaagcatt ggtcttccta ttttgatata acttcgtata
     2161 ggatacttta tacgaagtta tggggatcca attctttgcc aaaatgatga gacagcacaa
     2221 taaccagcac gttgcccagg agctgtagga aaaagaagaa ggcatgaaca tggttagcag
     2281 aggctctaga gccgccggtc acacgccaga agccgaaccc cgccctgccc cgtccccccc
     2341 gaaggcagcc gtccccccgc ggacagcccc gagccgctcc ccccgcacaa agggccctcc
     2401 cggagcccct caaggctttc acgcagccac agaaaagaaa caagccgtca ttaaaccaag
     2461 cgctaattac agcccggagg agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg
     2521 gtcagtcaga gccggggcgg gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg
     2581 cagcgcgcag cgactcccgc ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc
     2641 cgcctcgcca taaaaggaaa ctttcggagc gcgccgctct gattggctgc cgccgcacct
     2701 ctccgccycs ccccsccccs ccccycsccc csccccsccc csccyggcgc gcgccccccc
     2761 cccccccccc gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg
     2821 gggtggggag gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat
     2881 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     2941 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     3001 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     3061 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     3121 caatggtact cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3181 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3241 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3301 gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3361 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3421 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3481 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3541 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3601 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3661 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3721 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3781 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3841 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3901 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3961 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4021 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4081 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4141 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4201 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4261 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4321 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4381 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4441 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4501 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4561 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4621 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4681 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4741 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4801 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4861 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4921 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4981 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5041 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5101 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5161 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5221 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5281 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5341 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5401 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5461 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5521 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5581 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5641 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5701 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5761 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5821 ttaatgcag
//
