LOCUS       sequence-466203-        6174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2039m 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 6174)
  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 6174)
  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: AiP1175 - pAAV-AiE2039m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   152..292
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   163..292
                     /label=AAV2 ITR
     polyA_signal    332..808
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     590..609
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    840..1428
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             946..957
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1355..1375
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1453..2724,2725..2745,2746..2751))
                     /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      2748..2757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3599..3728
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3790..4378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3870..3889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4549..5340,5341..5409))
                     /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     5172..5191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5410..5514)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5582..5600
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5638..5660)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5760..5779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5796..6174,1..77))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5933..5954)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6145..6164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
       61 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
      121 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
      181 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      241 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      301 tcggtccgca cgtggttacc tacaaaatca gaaggacagg gaagggagca gtggttcacg
      361 cctgtaatcc cagcaatttg ggaggccaag gtgggtagat cacctgagat taggagttgg
      421 agaccagcct ggccaatatg gtgaaacccc gtctctacca aaaaaacaaa aattagctga
      481 gcctggtcat gcatgcctgg aatcccaaca actcgggagg ctgaggcagg agaatcgctt
      541 gaacccagga ggcggagatt gcagtgagcc aagattgtgc cactgcactc cagcttggtt
      601 cccaatagac cccgcaggcc ctacaggttg tcttcccaac ttgccccttg ctccatacca
      661 cccccctcca ccccataata ttatagaagg acacctagtc agacaaaatg atgcaactta
      721 attttattag gacaaggctg gtgggcactg gagtggcaac ttccagggcc aggagaggca
      781 ctggggaggg gtcacaggga tgccacccgt agatctctcg agcagcgctc ggtatcgatg
      841 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag
      901 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg
      961 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acataggcga gcagccaagg
     1021 aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc
     1081 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa
     1141 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc
     1201 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata
     1261 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
     1321 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
     1381 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt
     1441 gatatcgaat tctcagatcc gcctgttgat gtagctgctc aggtagtcca gcacctcctg
     1501 gctgatgatg ccgttccagg cggggtatct gatgctgccc tcggcgctgc ccttcagctg
     1561 ctcgatgtgc tgccactcct cgatggggtt ggtctcgtcc ttcagggcga tcatctcctt
     1621 gctgatgggg tcgtaggcgt agtacctgga caccagggcg aagtagtggt cggggatggc
     1681 ggtgatctgg tgggtgtagg tggtcctggc cacggcggag gccctcttgt cgctccagtt
     1741 gcccaccacg tttgtcagct cggtcaggcc cttcatgctc agaaagctgg tcatcaggtg
     1801 cctgccgatg tggctcttag ggccgttctt gatagcgaag atggggtagg gggcgttctt
     1861 cttcagggcc ttgttgtagc tgcgcaccag gttgtccttc agcagctggt actcctgctt
     1921 gttgctgctg ctgttgccgg tcctgttcac tctcttcagc acgggctcgc tgttcctcag
     1981 gaactcgtcc aggtacacca gggggtcgat cctgcctctg gcgctgaaaa agtagatgtg
     2041 cctggacacg cttgtcttgg tctcggtcac caggcactga atgatcacgc ccaggtactt
     2101 gttctgcacc agcttgaagc tcttggggtc cacgttcttg atgtcgctga acctgccgca
     2161 gttgatgaat gtggccagga acaggaactg gtacagggtc ttggtcttgg tgaacctgct
     2221 ggtgtactcg aagctgttca ggatcttctc ggtgatctcc cagatgctct cgccctcgga
     2281 cagcagggcc ttcagcatct tcttgctgtg gctgttgccc ttgtcggcct cctcgctgct
     2341 ctcgaactgc agctgcaggc tggacacgat gtcggtgatg tcgctctggt gcttctggcc
     2401 gttgtaaggg atgatggtga actcccaggc ggggatcagc ttcttcaggc tggcctccag
     2461 gatggtggcc ttctgggtct tgtacttgaa ctgcaggctc ttgttcacga tgtcgaagct
     2521 caggctgttg ctgatgatgg tgttgtagct catgaaggtg gccctcttga tggcggtgcc
     2581 gttgtgggtg atcatccagc acaggtaggt cagctcggcg gcacagctgg cgatcttctc
     2641 gccgctgggc ctctcgaatc tctccacgaa ctgccgcacc agcaccttgg ggggggtctt
     2701 gcacaggatg tcgaactggc tcatcacctt cctcttcttc ttaggagcca tggtggccac
     2761 gccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct
     2821 ccttaaggat agaggagaca agggggctgc attaccagac actagaggaa catgggtccc
     2881 taggccagag attccctatc atcgtgaccc tggctccagc actgataaag cagaggcggt
     2941 cccgtgggta gtgacagggt aagcggctga gccacaggca cagaggcgcc tctgggacag
     3001 atgggaggga ccctgagagt aggccctggg aagggggcca tggcgtcctg ccggcacacg
     3061 cggttgtccc tgtaggcctc tcttggaagg ggagctgctc ttcctccatc cttctccaag
     3121 tcagaagcaa ggctcacaat ggagcaatca agaccactct tcgggctttg ttctagtcaa
     3181 aggagacggg ggtgcttctg ggggctccat tcaaagccca actggtagga agccacagag
     3241 catctgaggc tgctctattt cagggcctct ctgcttcctg ttcccagctt cctgttctcc
     3301 ccccatcccc tgataccccg gaggggccgg gaggaccaaa tacaccagca aagaaagacc
     3361 tgggccgggg aggggcagct ccttgctcta accctctctc tctctctctc tctgcacggt
     3421 cccccagctc cgtgaaggat gtgcagtgtc ccaccctgct ctgagcggct cccctatgct
     3481 gtcgccagca cagctgtctg aggtcatgtg actgacaggc gtgaacgctt tgcagtggcc
     3541 tctgtctcat gggcaccaac cacttagtgc ccaaccagtc tttaacgcgt gcggccgcag
     3601 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3661 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3721 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4201 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4441 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4501 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4561 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4621 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4681 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4741 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4801 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4861 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4921 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4981 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5041 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5101 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5161 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5221 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5281 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5341 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5401 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5521 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5581 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5761 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     5821 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     5881 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     5941 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6001 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     6061 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6121 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaa
//
