LOCUS       sequence-466224-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2099m 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 6526)
  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 6526)
  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: AiP1248 - pAAV-AiE2099m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   374..503
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(565..1153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(645..664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1324..2115,2116..2184))
                     /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     1947..1966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2185..2289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2357..2375
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2413..2435)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2535..2554
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2571..3026)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2708..2729)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2920..2939
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3101..3241
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3112..3241
                     /label=AAV2 ITR
     polyA_signal    3281..3757
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3539..3558
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3789..4377
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3895..3906
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4304..4324
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4402..5673,5674..5694,5695..5700))
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      5697..5706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aatgagaaga agtagtgcct tgatcctata acatgcatac tgaagtacaa tccaactgtt
       61 actggttatc tatccacagc aaatagttaa ccactgtgag cctctgaaac cttgggatag
      121 taaatatttg ataagatcat taggaaaatt agttatacac agttctggag tccatggccc
      181 acagtagata tctcaggtat attttcccta atcacttatg aatctcactc acccagcttc
      241 accaattcct tcttctgctt gtaccttcct acatctactc tgtgatcaat tctaggatgc
      301 aaaattagat acagaaatgg aaaactagga catggtagaa cgtgctggaa ctgggtttga
      361 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
      421 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
      481 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      541 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      601 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      661 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      721 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      781 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      841 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      901 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      961 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1021 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1081 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1141 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1201 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1261 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1321 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1381 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1441 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1501 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1561 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1621 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1681 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1741 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1801 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1861 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1921 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1981 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2041 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2101 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2161 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2221 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2281 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     2341 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     2401 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2461 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2521 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
     2581 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2641 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
     2701 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2761 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
     2821 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2881 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2941 gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3001 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata
     3061 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct
     3121 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     3181 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     3241 tgcggccgct cggtccgcac gtggttacct acaaaatcag aaggacaggg aagggagcag
     3301 tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc acctgagatt
     3361 aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa aaaaacaaaa
     3421 attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc tgaggcagga
     3481 gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc actgcactcc
     3541 agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact tgccccttgc
     3601 tccataccac ccccctccac cccataatat tatagaagga cacctagtca gacaaaatga
     3661 tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact tccagggcca
     3721 ggagaggcac tggggagggg tcacagggat gccacccgta gatctctcga gcagcgctcg
     3781 gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag
     3841 acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag
     3901 ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cataggcgag
     3961 cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag
     4021 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg
     4081 gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt
     4141 gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa
     4201 ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga aagccatacg
     4261 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     4321 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattatc
     4381 gataagcttg atatcgaatt ctcagatccg cctgttgatg tagctgctca ggtagtccag
     4441 cacctcctgg ctgatgatgc cgttccaggc ggggtatctg atgctgccct cggcgctgcc
     4501 cttcagctgc tcgatgtgct gccactcctc gatggggttg gtctcgtcct tcagggcgat
     4561 catctccttg ctgatggggt cgtaggcgta gtacctggac accagggcga agtagtggtc
     4621 ggggatggcg gtgatctggt gggtgtaggt ggtcctggcc acggcggagg ccctcttgtc
     4681 gctccagttg cccaccacgt ttgtcagctc ggtcaggccc ttcatgctca gaaagctggt
     4741 catcaggtgc ctgccgatgt ggctcttagg gccgttcttg atagcgaaga tggggtaggg
     4801 ggcgttcttc ttcagggcct tgttgtagct gcgcaccagg ttgtccttca gcagctggta
     4861 ctcctgcttg ttgctgctgc tgttgccggt cctgttcact ctcttcagca cgggctcgct
     4921 gttcctcagg aactcgtcca ggtacaccag ggggtcgatc ctgcctctgg cgctgaaaaa
     4981 gtagatgtgc ctggacacgc ttgtcttggt ctcggtcacc aggcactgaa tgatcacgcc
     5041 caggtacttg ttctgcacca gcttgaagct cttggggtcc acgttcttga tgtcgctgaa
     5101 cctgccgcag ttgatgaatg tggccaggaa caggaactgg tacagggtct tggtcttggt
     5161 gaacctgctg gtgtactcga agctgttcag gatcttctcg gtgatctccc agatgctctc
     5221 gccctcggac agcagggcct tcagcatctt cttgctgtgg ctgttgccct tgtcggcctc
     5281 ctcgctgctc tcgaactgca gctgcaggct ggacacgatg tcggtgatgt cgctctggtg
     5341 cttctggccg ttgtaaggga tgatggtgaa ctcccaggcg gggatcagct tcttcaggct
     5401 ggcctccagg atggtggcct tctgggtctt gtacttgaac tgcaggctct tgttcacgat
     5461 gtcgaagctc aggctgttgc tgatgatggt gttgtagctc atgaaggtgg ccctcttgat
     5521 ggcggtgccg ttgtgggtga tcatccagca caggtaggtc agctcggcgg cacagctggc
     5581 gatcttctcg ccgctgggcc tctcgaatct ctccacgaac tgccgcacca gcaccttggg
     5641 gggggtcttg cacaggatgt cgaactggct catcaccttc ctcttcttct taggagccat
     5701 ggtggccacg ccagaagcaa atgtaagcaa tagatggctc tgccctgact tttatgccca
     5761 gcccgagctc gcttcgtata aaccataact caaattctgt gtgttgaccc tgaatgacca
     5821 taacatatat tatccaagta tctgtgtcta ttaattggta ttataatgtt taacataata
     5881 aggacagatt taatcattta agttctgttc caattaagtc ctaagtttat caatgggctt
     5941 cactcatttt agtggcttca ataaaacaaa taacccaaca atacagatat caatagatgt
     6001 ttttgcttct tcttagaacc tcttgcagat gcttggagcc agtgattctg agtaagatga
     6061 catcagagca gatatgaaac aataaaatca ggtccaagaa tcccagctga aaatttaaaa
     6121 aggaatgctc cgccttgcat ttcacaagtc gtaaaatttc cccttcagta cccatgcaag
     6181 ttggctgggg aaagaagggg ccaggcatga ttccctgaga aatcagtgaa tatttatctc
     6241 tctccagctt catgtaacca tggcaggtct tgagatgctt gaaaatcttg ccaaatatta
     6301 ctaaaccctt actctaatag agttctggtg ctatatcttt aataactcac taactattcc
     6361 aagcatttta atatgctctt gttttgcttt cagccaggct ccaaaataga gcatctgtta
     6421 ttttatagat gggaaattaa agcctgaagc ccagaatgct tggctgagta actaacaaga
     6481 ggctgagctg gaatgcattg aagcatacat ccagggagga tgggac
//
