LOCUS       sequence-466216-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0079h 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 6013)
  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 6013)
  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: AiP1230 - pAAV-AiE0079h-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   118..247
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(309..897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(389..408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1068..1859,1860..1928))
                     /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     1691..1710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1929..2033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2101..2119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2157..2179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2279..2298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2315..2770)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2452..2473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2664..2683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2845..2985
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2856..2985
                     /label=AAV2 ITR
     polyA_signal    3025..3501
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3283..3302
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3533..4121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3639..3650
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4048..4068
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4146..5417,5418..5438,5439..5444))
                     /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      5441..5450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agagtacaaa attaaaagag gcactcactt atgtaagtgc atttgcagga ctctgaaagt
       61 gaatgcctct ttaaatttca tatcctgggt atcttattca cctacgcgtg cggccgcagg
      121 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
      181 ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
      241 ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      481 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      721 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2041 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     2101 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     2161 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     2221 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     2281 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
     2341 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
     2401 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
     2461 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2521 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     2581 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2641 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     2701 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2761 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
     2821 ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg
     2881 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2941 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc
     3001 gcacgtggtt acctacaaaa tcagaaggac agggaaggga gcagtggttc acgcctgtaa
     3061 tcccagcaat ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag
     3121 cctggccaat atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt
     3181 catgcatgcc tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca
     3241 ggaggcggag attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata
     3301 gaccccgcag gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct
     3361 ccaccccata atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat
     3421 taggacaagg ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga
     3481 ggggtcacag ggatgccacc cgtagatctc tcgagcagcg ctcggtatcg atgcggggag
     3541 gcggcccaaa gggagatccg actcgtctga gggcgaaggc gaagacgcgg aagaggccgc
     3601 agagccggca gcaggccgcg ggaaggaagg tccgctggat tgagggccga agggacgtag
     3661 cagaaggacg tcccgcgcag aatccaggtg gcaacatagg cgagcagcca aggaaaggac
     3721 gatgatttcc ccgacaacac cacggaattg tcagtgccca acagccgagc ccctgtccag
     3781 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa tagggagggg gaaagcgaaa
     3841 gtcccggaaa ggagctgaca ggtggtggca atgccccaac cagtgggggt tgcgtcagca
     3901 aacacagtgc acaccacgcc acgttgcctg acaacgggcc acaactcctc ataaagagac
     3961 agcaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     4021 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     4081 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatcgataag cttgatatcg
     4141 aattctcaga tccgcctgtt gatgtagctg ctcaggtagt ccagcacctc ctggctgatg
     4201 atgccgttcc aggcggggta tctgatgctg ccctcggcgc tgcccttcag ctgctcgatg
     4261 tgctgccact cctcgatggg gttggtctcg tccttcaggg cgatcatctc cttgctgatg
     4321 gggtcgtagg cgtagtacct ggacaccagg gcgaagtagt ggtcggggat ggcggtgatc
     4381 tggtgggtgt aggtggtcct ggccacggcg gaggccctct tgtcgctcca gttgcccacc
     4441 acgtttgtca gctcggtcag gcccttcatg ctcagaaagc tggtcatcag gtgcctgccg
     4501 atgtggctct tagggccgtt cttgatagcg aagatggggt agggggcgtt cttcttcagg
     4561 gccttgttgt agctgcgcac caggttgtcc ttcagcagct ggtactcctg cttgttgctg
     4621 ctgctgttgc cggtcctgtt cactctcttc agcacgggct cgctgttcct caggaactcg
     4681 tccaggtaca ccagggggtc gatcctgcct ctggcgctga aaaagtagat gtgcctggac
     4741 acgcttgtct tggtctcggt caccaggcac tgaatgatca cgcccaggta cttgttctgc
     4801 accagcttga agctcttggg gtccacgttc ttgatgtcgc tgaacctgcc gcagttgatg
     4861 aatgtggcca ggaacaggaa ctggtacagg gtcttggtct tggtgaacct gctggtgtac
     4921 tcgaagctgt tcaggatctt ctcggtgatc tcccagatgc tctcgccctc ggacagcagg
     4981 gccttcagca tcttcttgct gtggctgttg cccttgtcgg cctcctcgct gctctcgaac
     5041 tgcagctgca ggctggacac gatgtcggtg atgtcgctct ggtgcttctg gccgttgtaa
     5101 gggatgatgg tgaactccca ggcggggatc agcttcttca ggctggcctc caggatggtg
     5161 gccttctggg tcttgtactt gaactgcagg ctcttgttca cgatgtcgaa gctcaggctg
     5221 ttgctgatga tggtgttgta gctcatgaag gtggccctct tgatggcggt gccgttgtgg
     5281 gtgatcatcc agcacaggta ggtcagctcg gcggcacagc tggcgatctt ctcgccgctg
     5341 ggcctctcga atctctccac gaactgccgc accagcacct tggggggggt cttgcacagg
     5401 atgtcgaact ggctcatcac cttcctcttc ttcttaggag ccatggtggc cacgccagaa
     5461 gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga gctcaatctc
     5521 accaatcacc actaaagaac ttactcatgt aaccaaatac ccctgttccc caaaaaccta
     5581 tggaaattaa aaataaattt aaaaaataaa aaaattttag tggcttaaaa cagcacccat
     5641 ttattatctc acagctgtgt aggtcaaaat ttccaattcc cataaatacc agctgcataa
     5701 aagacttcct gtttatgggt acggtgtggt tcagtggagt cctctgctta gagtgccaca
     5761 gtattgaaat agaagtgcca ggcaggctgt attcctttct gggagttcta agaatgaatc
     5821 cacttccaat atcactcttg ctattggtag aattcagttc catgtggtta ttctactgag
     5881 gtcctcattt ccttgctggc tgtcagctca ggatatatag ctttcggagg ccacccacaa
     5941 tctttggctc atggctcgtt ttctctgcct tcaaagctag cagtggacga gaatgagatg
     6001 aagtgagtca att
//
