LOCUS       sequence-459312-        5852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre in excitatory neurons; Cre expression is attenuated by
            PEST sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  AUTHORS   Drexler R, Drinnenberg A, Gavish A, Yalcin B, Shamardani K, Rogers 
            AE, Mancusi R, Trivedi V, Taylor KR, Kim YS, Woo PJ, Soni N, Su M, 
            Ravel A, Tatlock E, Midler A, Wu SH, Ramakrishnan C, Chen R, 
            Ayala-Sarmiento AE, Fernandez Pacheco DR, Siverts L, Daigle TL, 
            Tasic B, Zeng H, Breunig JJ, Deisseroth K, Monje M
  TITLE     Cholinergic neuronal activity promotes diffuse midline glioma growth
            through muscarinic signaling.
  JOURNAL   Cell. 2025 Aug 21;188(17):4640-4657.e30. doi: 
            10.1016/j.cell.2025.05.031. Epub 2025 Jun 19.
  PUBMED    40541184
REFERENCE   2  (bases 1 to 5852)
  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.2025.05.031"; journalName: "Cell"; date: 
            "2025-08-21- 21"; volume: "188"; issue: "17"; pages: "4640-4657.e30"
COMMENT     Sequence Label: pAAV-eHGT_78h-Cre_PEST
FEATURES             Location/Qualifiers
     source          1..5852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1437..1457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2629..3217
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2682..2702)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3100..3111)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3249..3725
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3448..3467)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3765..3905
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3765..3894
                     /label=AAV2 ITR
     rep_origin      3980..4435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4277..4298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4452..4471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4571..4593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4631..4649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtgaaatc ataaatgctg agggtagtct gcctcaggta cacactgaga
      841 aactgcttta atgtaacctg acccacggtt attagtgaaa atatcacttt tgttgttacc
      901 ttattcccaa caaattcatt tctgctttaa tggaaaagat ccgggttcac actaatcagg
      961 cccaacggaa ggccatatta gcaatttggc aggtacccga gggccatacc taatctgcat
     1021 aaaatgaagc agattgcaac cgccctcatc ttttttattt ttaaactggt ttttgaagca
     1081 gagcataaaa tctcagaggg agagacagaa gatgctagtg catacatttt ccttcatgcc
     1141 tttattttca ttctttttgc acaaaccatc ttcctgaatg gctgtttacc taaagaagaa
     1201 taacaaaata aaaggtgcta ggaaatggag taggcagaga tcacaaatgt ttaattaaaa
     1261 aaaaaaaaag tcatgtactt tcatagatat tcacaatcct ctctagtata ctttcaaatc
     1321 agttttaatt tcagtttagt gtttttatgt tttgtgaaga tacgcgagct cgggctgggc
     1381 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc accatgccca
     1441 agaagaagag gaaggtgtcc aatctcctga ctgttcacca gaacctccct gcgctgccag
     1501 tagatgccac tagcgatgag gtcaggaaaa atctcatgga tatgtttagg gatagacagg
     1561 cgttttctga acacacctgg aaaatgctgc ttagcgtgtg ccgatcctgg gcagcctggt
     1621 gtaagctgaa caatcgcaaa tggttccccg ccgagccgga ggacgtgcgc gattacctgc
     1681 tgtatctcca ggcaagaggg ctggctgtca agactatcca gcagcacttg ggccaactga
     1741 atatgctgca tcgacgcagc gggctccccc ggcctagcga ttcaaacgca gtctcccttg
     1801 ttatgaggag aattagaaag gaaaacgtag atgcgggtga gagggctaag caggctctcg
     1861 cttttgagcg gactgatttc gaccaggtca gatccctgat ggagaacagc gatcggtgcc
     1921 aggacatcag gaacctcgca tttctgggaa ttgcatataa cacacttctg cgcatagctg
     1981 agatcgcccg gatcagagtg aaagacatca gtcgaacgga cggcggccgg atgcttattc
     2041 atattggacg cacaaagaca ttggtcagca ccgctggcgt tgaaaaggcc ttgtccctgg
     2101 gcgtaacgaa gctggtggaa agatggatct cagtgtccgg cgtggctgac gaccctaata
     2161 attacttgtt ctgtcgagtg agaaaaaacg gagtcgccgc gccctctgcc accagccaat
     2221 tgagtacacg ggcccttgaa gggatctttg aggcaaccca ccgactcata tacggagcca
     2281 aggatgacag tggccagagg tatctcgcct ggtcaggtca ttctgctagg gtgggggccg
     2341 cacgagacat ggcgcgggca ggagtctcca taccagagat tatgcaagct ggaggttgga
     2401 caaatgtgaa catcgttatg aactatatcc gcaatcttga ctctgaaacc ggggccatgg
     2461 tgagactgct cgaagatggt gacagccatg gcttcccgcc ggaggtggag gagcaggatg
     2521 atggcacgct gcccatgtct tgtgcccagg agagcgggat ggaccgtcac cctgcagcct
     2581 gtgcttctgc taggatcaat gtgtgaattc gatatcaagc ttatcgataa tcaacctctg
     2641 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2701 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2761 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2821 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2881 gccaccacct gtcagctcct ttccggaact ttcgctttcc ccctccctat tgccacggcg
     2941 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3001 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3061 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3121 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3181 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3241 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3301 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3361 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3421 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3481 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3541 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3601 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3661 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3721 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3781 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3841 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3901 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3961 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4021 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4201 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4261 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4321 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4381 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4441 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4501 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4561 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4621 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4681 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tc
//
