LOCUS       sequence-461956-        6010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2035m 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 6010)
  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 6010)
  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: AiP1172 - pAAV-AiE2035m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             complement(371..382)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(814..819,820..840,841..2112)
                     /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    2137..2725
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2190..2210)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2608..2619)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2757..3233
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2956..2975)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3273..3413
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3273..3402
                     /label=AAV2 ITR
     rep_origin      3488..3943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3575..3594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3785..3806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3960..3979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4079..4101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4139..4157)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4225..4329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4330..4398,4399..5190)
                     /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(4548..4567)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5361..5949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5850..5869
                     /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 gcgtgtagct atgcttccct agccgttcac ataactttct
      181 tattttactt gaatttcttt agtaaagttt tgatacgaac aaccgccaac ttaccaactc
      241 aacaattact catgctgagg agagccttga gtttctggag acagagcagt gaaccaagag
      301 acaaatacat taataaatat ttattgttcc agggattttt ctagacatgc agatggagaa
      361 ttgtgctgtc tctgccctca atgggtagat gaatagatga aagcagtata agatacacgg
      421 gctcataaat agcagatgag acatgcaaat cagagccaag agggaagggt ttcagagaaa
      481 agacagaaaa gaatgactga cttgtgttat cctgggtaga ccaattacca agccctgtat
      541 acaggcttga ttgtgaaatt ctgcccaaag tatgcagaaa aatgctgatt aaattgggca
      601 actggtgaag actttggata ttaaattttc aataatcgag tgagtcatga aaacatgctt
      661 ttatggtcat ttacattggg gatgttgatt tccaatgaca ctctacgggc tcaattaagc
      721 agggggataa ttcagtactt aaggagctcg ggctgggcat aaaagtcagg gcagagccat
      781 ctattgctta catttgcttc tggcgtggcc accatggctc ctaagaagaa gaggaaggtg
      841 atgagccagt tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg
      901 gagagattcg agaggcccag cggcgagaag atcgccagct gtgccgccga gctgacctac
      961 ctgtgctgga tgatcaccca caacggcacc gccatcaaga gggccacctt catgagctac
     1021 aacaccatca tcagcaacag cctgagcttc gacatcgtga acaagagcct gcagttcaag
     1081 tacaagaccc agaaggccac catcctggag gccagcctga agaagctgat ccccgcctgg
     1141 gagttcacca tcatccctta caacggccag aagcaccaga gcgacatcac cgacatcgtg
     1201 tccagcctgc agctgcagtt cgagagcagc gaggaggccg acaagggcaa cagccacagc
     1261 aagaagatgc tgaaggccct gctgtccgag ggcgagagca tctgggagat caccgagaag
     1321 atcctgaaca gcttcgagta caccagcagg ttcaccaaga ccaagaccct gtaccagttc
     1381 ctgttcctgg ccacattcat caactgcggc aggttcagcg acatcaagaa cgtggacccc
     1441 aagagcttca agctggtgca gaacaagtac ctgggcgtga tcattcagtg cctggtgacc
     1501 gagaccaaga caagcgtgtc caggcacatc tactttttca gcgccagagg caggatcgac
     1561 cccctggtgt acctggacga gttcctgagg aacagcgagc ccgtgctgaa gagagtgaac
     1621 aggaccggca acagcagcag caacaagcag gagtaccagc tgctgaagga caacctggtg
     1681 cgcagctaca acaaggccct gaagaagaac gccccctacc ccatcttcgc tatcaagaac
     1741 ggccctaaga gccacatcgg caggcacctg atgaccagct ttctgagcat gaagggcctg
     1801 accgagctga caaacgtggt gggcaactgg agcgacaaga gggcctccgc cgtggccagg
     1861 accacctaca cccaccagat caccgccatc cccgaccact acttcgccct ggtgtccagg
     1921 tactacgcct acgaccccat cagcaaggag atgatcgccc tgaaggacga gaccaacccc
     1981 atcgaggagt ggcagcacat cgagcagctg aagggcagcg ccgagggcag catcagatac
     2041 cccgcctgga acggcatcat cagccaggag gtgctggact acctgagcag ctacatcaac
     2101 aggcggatct gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2161 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2221 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2281 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2341 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2401 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2461 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2521 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     2581 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2641 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2701 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     2761 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2821 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2881 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2941 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3001 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3061 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3121 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3181 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3241 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3301 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3361 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3481 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3541 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3781 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3841 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3901 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3961 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4141 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4201 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4261 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4321 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4381 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4441 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4501 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4561 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4621 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4681 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4741 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4801 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4861 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4921 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4981 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5041 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5101 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5161 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5221 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5281 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5341 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5401 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5461 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5521 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5581 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5641 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5701 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5761 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5821 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5881 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5941 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6001 gctcacatgt
//
