LOCUS       sequence-466298-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2392m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4757)
  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"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP1736 - pAAV-AiE2392m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   238..378
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   249..378
                     /label=AAV2 ITR
     polyA_signal    496..607
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     794..814
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(907..1617,1618..1620,1621..1623))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(949..970)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1298..1317
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1557..1578
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1620..1629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2268..2397
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2459..3047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2539..2558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3218..4009,4010..4078))
                     /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     3841..3860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4079..4183)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4251..4269
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4307..4329)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4429..4448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4465..4757,1..163))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4602..4623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
       61 aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta
      121 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
      181 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct
      241 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
      301 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      361 ccatcactag gggttcctgc ggccgctcgg tccgcacgtg agatctcatg cctgctattg
      421 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
      481 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
      541 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
      601 ggcacagtcg aagatctctc gagccacgga attgtcagtg cccaacagcc gagcccctgt
      661 ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta accaggattt
      721 atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa
      781 agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc
      841 acaaattttg taatccagag gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc
      901 gacttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
      961 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact ggtagctcag
     1021 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1081 gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat
     1141 gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag
     1201 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1261 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1321 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1381 ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccgtagccca gggtggtcac
     1441 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt
     1501 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     1561 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     1621 catggtggcg accggtagcg ctagcttcga aagatctgga tcccagaagc aaatgtaagc
     1681 aatagatggc tctgccctga cttttatgcc cagcccgagc tccgtttttc atctgcattg
     1741 aaccttagtt tcttatttcc ccaaatcata cctattcact caaaagtctt tacaagtgtt
     1801 tcccatagaa gtagtattct taagagagtc tctcagacat ctgagcttaa tggattttgc
     1861 tttaaagttc tatgctttca ttctttcttt tcccttgcta aaccttttta aacaagtgag
     1921 ttgaataaaa caatttttcc ccctcagagg ttacaccatg cttatttcac agcccgattt
     1981 agtcaagctt tttggctttt aactcaaagg aatgacattt taaaaatgtt ctcttatggg
     2041 tatatttctg ttgatagagt tctgaacttt aaaaaaaata gatgcaagaa tcacatttaa
     2101 gataaagaaa tgtgcatagt tcaactttac tactaagcaa tcttagaaaa atgatccata
     2161 atctacatta atgtgtagtt cattcacaaa atgagtgcag tatttatgct cttcataaaa
     2221 ccattggaaa agttctcgat tattactctt acaacgcgtg cggccgcagg aacccctagt
     2281 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2341 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca
     2401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2641 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2881 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3181 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3241 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3301 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3361 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3421 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3481 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3541 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3601 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3661 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3721 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3781 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3841 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3901 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3961 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4021 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4081 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4141 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4201 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4261 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4321 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4381 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4441 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     4501 taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt
     4561 ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc
     4621 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4681 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4741 taaatcggaa ccctaaa
//
