LOCUS       sequence-466225-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2101m 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 6534)
  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 6534)
  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: AiP1250 - pAAV-AiE2101m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      698..707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(704..709,710..730,731..2002)
                     /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"
     misc_feature    2027..2615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2080..2100)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2498..2509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2647..3123
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2846..2865)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3163..3303
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3163..3292
                     /label=AAV2 ITR
     rep_origin      3378..3833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3675..3696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3850..3869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3969..3991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4029..4047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4115..4219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4220..4288,4289..5080)
                     /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(4438..4457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5251..5839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5740..5759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5901..6030
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtcatggag ctgtcccaag atggaacaca tatttggatt caataagaaa gcccccacat
       61 cattttgagt atgtttgcaa agcaaaaata gctgctgcta tgtgtttcat ttttgccaaa
      121 attattacat tcatcccaga agagaaagcg acgtgaacct cttctgggaa tgatttcggc
      181 agccacaggc cagggagtga aactacctgg gaatgttcat tttctctagc agatggctcc
      241 tgcttgttag aagggttttc ctagaccatg cccttgaatt tggataaatt gggaaaaaaa
      301 ttaaacattt ttggatttga ccatgaatta gtttttctct ttttatctgt cttgtatttg
      361 tttagctgac atatttcttg ggttggtttc ttttatagga agcaaacttt gaaggtgtgt
      421 caacaaactc caaaattggt agaataaaga ccccttcttt tccaactcct ctagctgatt
      481 ttttttatgc tttttttttt ccttgtagaa tagtgggatg actgtcaatg gagtactcag
      541 agaataagta aggccttccc aggagtatgt gtttccagga gcactggaaa gatcaattcc
      601 ttgcatcaaa gcatttctgc attcataaca cctgagctcg ggctgggcat aaaagtcagg
      661 gcagagccat ctattgctta catttgcttc tggcgtggcc accatggctc ctaagaagaa
      721 gaggaaggtg atgagccagt tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg
      781 gcagttcgtg gagagattcg agaggcccag cggcgagaag atcgccagct gtgccgccga
      841 gctgacctac ctgtgctgga tgatcaccca caacggcacc gccatcaaga gggccacctt
      901 catgagctac aacaccatca tcagcaacag cctgagcttc gacatcgtga acaagagcct
      961 gcagttcaag tacaagaccc agaaggccac catcctggag gccagcctga agaagctgat
     1021 ccccgcctgg gagttcacca tcatccctta caacggccag aagcaccaga gcgacatcac
     1081 cgacatcgtg tccagcctgc agctgcagtt cgagagcagc gaggaggccg acaagggcaa
     1141 cagccacagc aagaagatgc tgaaggccct gctgtccgag ggcgagagca tctgggagat
     1201 caccgagaag atcctgaaca gcttcgagta caccagcagg ttcaccaaga ccaagaccct
     1261 gtaccagttc ctgttcctgg ccacattcat caactgcggc aggttcagcg acatcaagaa
     1321 cgtggacccc aagagcttca agctggtgca gaacaagtac ctgggcgtga tcattcagtg
     1381 cctggtgacc gagaccaaga caagcgtgtc caggcacatc tactttttca gcgccagagg
     1441 caggatcgac cccctggtgt acctggacga gttcctgagg aacagcgagc ccgtgctgaa
     1501 gagagtgaac aggaccggca acagcagcag caacaagcag gagtaccagc tgctgaagga
     1561 caacctggtg cgcagctaca acaaggccct gaagaagaac gccccctacc ccatcttcgc
     1621 tatcaagaac ggccctaaga gccacatcgg caggcacctg atgaccagct ttctgagcat
     1681 gaagggcctg accgagctga caaacgtggt gggcaactgg agcgacaaga gggcctccgc
     1741 cgtggccagg accacctaca cccaccagat caccgccatc cccgaccact acttcgccct
     1801 ggtgtccagg tactacgcct acgaccccat cagcaaggag atgatcgccc tgaaggacga
     1861 gaccaacccc atcgaggagt ggcagcacat cgagcagctg aagggcagcg ccgagggcag
     1921 catcagatac cccgcctgga acggcatcat cagccaggag gtgctggact acctgagcag
     1981 ctacatcaac aggcggatct gagaattcga tatcaagctt atcgataatc aacctctgga
     2041 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2101 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2161 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2221 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2281 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2341 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2401 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     2461 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2521 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2581 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     2641 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2701 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2761 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2821 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2881 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     2941 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     3001 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     3061 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3121 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3181 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3241 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3301 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3361 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3721 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4021 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4081 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5701 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5761 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5821 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5881 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     5941 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     6001 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtcattgg gtgaggaatt
     6061 gcaaaaccac tgcagtcaat ttgtaagttt aaaaagtgtt agtttgtgga gcataaaagt
     6121 aaaaaaaaaa aaaaatcaat ctactctgac tcaaaagtct cttctattct ctatagctga
     6181 aacaaaaaaa ggttacaaag tcattcagaa gttgctacct tacacaagtg ctttgcttag
     6241 tggagcagga ttctatgcgt atggcagagg gagggacgca gatctccaca ttgaacacaa
     6301 tctgcagtaa tgggaaagcc tgctgtaaaa taacacccaa catgttagaa cgtgttcgac
     6361 cggctgaaat tggaatggag atttgtaaac atatttatag agttgcagac tacacaatgc
     6421 agtcagaata ttcagagtta ctatttaaat ctgctgggat attgttctta tttttacgtt
     6481 cctccaaaat aggtttcagt agctttcgta ggtttgtgga gagaaagtta actc
//
