LOCUS       sequence-466218-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2065m 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 6231)
  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 6231)
  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: AiP1237 - pAAV-AiE2065m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     474..493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   635..764
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1663..1672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1669..1674,1675..1695,1696..2967)
                     /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    2992..3580
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3045..3065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3463..3474)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3612..4088
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4128..4268
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4128..4257
                     /label=AAV2 ITR
     rep_origin      4343..4798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4430..4449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4640..4661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4815..4834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4934..4956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4994..5012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5080..5184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5185..5253,5254..6045)
                     /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(5403..5422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6216..6231,1..573)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
       61 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      121 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      181 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      241 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      301 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      361 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      421 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      481 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      541 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      601 ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg
      661 ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      721 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtcg
      781 cttcttaaac ttgcagagag ctcacggtct gcctgtgctg ctaccattct tattgaataa
      841 cccactggta caacctgtaa gactatgtca gatttgcatg acccatgcaa atagaaacat
      901 ttagtttcac cttgaaagat atataggatt cattttgttt taaaaattct atcaagcact
      961 tggggtttat gttcagcacc agtagtcgtt ccatctccta ctggtactcc gtgatatgag
     1021 catgcttatg gcctaaggct actccttgag aagataattt gttaggctac aggccacgat
     1081 tggctctctg ctgtccttcc caaggaaaga cagcacacat ccactcccca catacaggcc
     1141 gtatatttca caaagtatta tgtgaactgt tgtacaagac tggattgcag gactggtcag
     1201 tccatgctta tataacagct cagctgaaga caattgtttt cagtaatttc cactgagctc
     1261 agttctccaa ggagtcacaa tgctttctgg ttgcatgcag ggcacttttg gaagctgcag
     1321 tcaattctgg aggccaccag ggcaccatca gcacgtagca gcaattactg actaagtggc
     1381 actctggttc cattcttctg aggggcctac ttagaggcaa ggcgaagttg ctctagtgaa
     1441 actgaacaaa atacctaatt tctagagact gtaagaaaag tttgaaatgc cttgtgaatg
     1501 tgttgacttg acaacaaaat acatactaaa aatcaagttt ctggtccaaa ctcagcccaa
     1561 agaaaaagtt agagaaccag atgatgttat gtcttaagga gctcgggctg ggcataaaag
     1621 tcagggcaga gccatctatt gcttacattt gcttctggcg tggccaccat ggctcctaag
     1681 aagaagagga aggtgatgag ccagttcgac atcctgtgca agaccccccc caaggtgctg
     1741 gtgcggcagt tcgtggagag attcgagagg cccagcggcg agaagatcgc cagctgtgcc
     1801 gccgagctga cctacctgtg ctggatgatc acccacaacg gcaccgccat caagagggcc
     1861 accttcatga gctacaacac catcatcagc aacagcctga gcttcgacat cgtgaacaag
     1921 agcctgcagt tcaagtacaa gacccagaag gccaccatcc tggaggccag cctgaagaag
     1981 ctgatccccg cctgggagtt caccatcatc ccttacaacg gccagaagca ccagagcgac
     2041 atcaccgaca tcgtgtccag cctgcagctg cagttcgaga gcagcgagga ggccgacaag
     2101 ggcaacagcc acagcaagaa gatgctgaag gccctgctgt ccgagggcga gagcatctgg
     2161 gagatcaccg agaagatcct gaacagcttc gagtacacca gcaggttcac caagaccaag
     2221 accctgtacc agttcctgtt cctggccaca ttcatcaact gcggcaggtt cagcgacatc
     2281 aagaacgtgg accccaagag cttcaagctg gtgcagaaca agtacctggg cgtgatcatt
     2341 cagtgcctgg tgaccgagac caagacaagc gtgtccaggc acatctactt tttcagcgcc
     2401 agaggcagga tcgaccccct ggtgtacctg gacgagttcc tgaggaacag cgagcccgtg
     2461 ctgaagagag tgaacaggac cggcaacagc agcagcaaca agcaggagta ccagctgctg
     2521 aaggacaacc tggtgcgcag ctacaacaag gccctgaaga agaacgcccc ctaccccatc
     2581 ttcgctatca agaacggccc taagagccac atcggcaggc acctgatgac cagctttctg
     2641 agcatgaagg gcctgaccga gctgacaaac gtggtgggca actggagcga caagagggcc
     2701 tccgccgtgg ccaggaccac ctacacccac cagatcaccg ccatccccga ccactacttc
     2761 gccctggtgt ccaggtacta cgcctacgac cccatcagca aggagatgat cgccctgaag
     2821 gacgagacca accccatcga ggagtggcag cacatcgagc agctgaaggg cagcgccgag
     2881 ggcagcatca gataccccgc ctggaacggc atcatcagcc aggaggtgct ggactacctg
     2941 agcagctaca tcaacaggcg gatctgagaa ttcgatatca agcttatcga taatcaacct
     3001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3121 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3181 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3241 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3301 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3361 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3421 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3481 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3541 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3601 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3661 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3721 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3781 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3841 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3901 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3961 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     4021 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     4081 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4141 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4201 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4261 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4321 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4381 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4441 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4501 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4561 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4621 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4681 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4741 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4801 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4861 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4921 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4981 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5041 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5401 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5701 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt t
//
