LOCUS       sequence-471061-        5079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0086m 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 5079)
  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 5079)
  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: AiP20171 -
            pAAV-hsA2-AiE0086m-minRho-iCre(R297...
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1015..1024
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1027..1047
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2113..2224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2342..2482
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2342..2471
                     /label=AAV2 ITR
     rep_origin      2557..3012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2644..2663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2854..2875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3029..3048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3148..3170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3208..3226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3294..3398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3399..3467,3468..4259)
                     /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(3617..3636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4430..5018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4919..4938
                     /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 gcgtttagaa caatggctgg cccatagtaa atgccgtgtt
      181 agtgtgttag ttgctgttct tccacgtcag aagaggcaca gacaaattac caccaggtgg
      241 cgctcagagt ctgcggaggc atcacaacag ccctgaattt gaatcctgct ctgccactgc
      301 ctagttgaga ccttttacta cctgactagc tgtttgtgta ttttaggtgt ttgtttcact
      361 ctcatacccc agcctcattt caaagctagg gatcatagcg ccagccttca gaaaagagac
      421 tcaaaaggag caaaaacgaa gagtgcaggg ctgctagata gactccatcc agggaccgag
      481 cctgtgacga atccagcccc tggcaagagc ccgacaggca ggtgtaatca gcaaatttct
      541 ccggagacat cagagcagat ccctcagcaa gcacagcttc taaatttgtg ctcaagaatg
      601 attaatctgt ttaagcaaat tccttgagca taattaagat gccctaatca aggaagaaga
      661 ttattatgcg gcagccttag gcactgattt ggtggcaaga gcccgtgacc acaaggcctc
      721 aataacttgg tggggtagtg aaggtgacag agtggcttag gctgcaggga ggtgactggg
      781 gtgggggtgg ggggaggtta ggccactggg aagccgagaa tatttaagga atgggaattc
      841 gatatcataa tcaaccatag gtaccgagct cgggattcag ccgggagctt agggagggga
      901 ggtcacttca taagggcctg gggggggagt tggagccacg agtcgtccag ccggagcccc
      961 gtgtggctga gctccggcct cagaagcatc cccgggttgg atccttcgaa gctagccacc
     1021 atggtgccca agaagaagag gaaagtctcc aacctgctga ctgtgcacca aaacctgcct
     1081 gccctccctg tggatgccac ctctgatgaa gtcaggaaga acctgatgga catgttcagg
     1141 gacaggcagg ccttctctga acacacctgg aagatgctcc tgtctgtgtg cagatcctgg
     1201 gctgcctggt gcaagctgaa caacaggaaa tggttccctg ctgaacctga ggatgtgagg
     1261 gactacctcc tgtacctgca agccagaggc ctggctgtga agaccatcca acagcacctg
     1321 ggccagctca acatgctgca caggagatct ggcctgcctc gcccttctga ctccaatgct
     1381 gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg atgctgggga gagagccaag
     1441 caggccctgg cctttgaacg cactgacttt gaccaagtca gatccctgat ggagaactct
     1501 gacagatgcc aggacatcag gaacctggcc ttcctgggca ttgcctacaa caccctgctg
     1561 cgcattgccg aaattgccag aatcagagtg aaggacatct cccgcaccga tggtgggaga
     1621 atgctgatcc acattggcag gaccaagacc ctggtgtcca cagctggtgt ggagaaggcc
     1681 ctgtccctgg gggttaccaa gctggtggag agatggatct ctgtgtctgg tgtggctgat
     1741 gaccccaaca actacctgtt ctgccgggtc agaaagaatg gtgtggctgc cccttctgcc
     1801 acctcccaac tgtccacccg ggccctggaa gggatctttg aggccaccca ccgcctgatc
     1861 tatggtgcca aggatgactc tgggcagaga tacctggcct ggtctggcca ctctgccaga
     1921 gtgggtgctg ccaccgacat ggccagggct ggtgtgtcca tccctgaaat catgcaggct
     1981 ggtggctgga ccaatgtgaa cattgtgatg aactacatca gaaacctgga ctctgagact
     2041 ggggccatgg tgaggctgct cgaagatggg gactgaggcg cgccgaattc aagcttctcg
     2101 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2161 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2221 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2281 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     2341 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2401 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2461 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2521 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2581 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2641 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2701 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2761 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2821 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2881 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     2941 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3001 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3061 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3121 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3181 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3241 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3301 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3361 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3421 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3481 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3541 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3601 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3661 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3721 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3781 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3841 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3901 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3961 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4021 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4081 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4141 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4201 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4261 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4321 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4381 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4441 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4501 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4561 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4621 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4681 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4741 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4801 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4861 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4921 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4981 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5041 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
