LOCUS       sequence-474095-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0330h 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 6106)
  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 6106)
  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: AiP12783 - pAAV-AiE0330h-minBG-FlpO-WPRE3-HGHp...
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     RBS             809..817
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      904..913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(910..915,916..936,937..2208)
                     /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    2233..2821
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2286..2306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2704..2715)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2853..3329
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3369..3509
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3369..3498
                     /label=AAV2 ITR
     rep_origin      3584..4039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3881..3902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4056..4075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4175..4197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4235..4253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4321..4425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4426..4494,4495..5286)
                     /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(4644..4663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5457..6045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5946..5965
                     /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 gcgtttgtgg aggctgacaa gtctcaatac ctacagtcgg
      181 caagctagag acccaggaga acctatggtg tagtccaggc tcattccaaa ggcctaagaa
      241 ccaagcgagc caatgatgta gttctagtgt gaaaacctgc aggctctaga cacaggaaga
      301 agccaatgtt tcagttaaag tccaagggca ggaaacacca aggtcccagc tcaaacatag
      361 tgagacagaa ggaattctct cttactcagc cttttgttct attcaggctt tcaacagatt
      421 ggatgaggcc cacccacaat ctgctttacc cagtctactg atttagatgt taatctcatc
      481 cagaaacacc ctcacagaca cacctagaat aatgtttgac caaatgtctg ggcaccctgt
      541 tacccagtca agtggatgca taaaattaaa caccataaag actaaagcct gtagatgaat
      601 atctggtaac acctcacaga ggagagacct aaagctgtga taacgtctct gaagccaaga
      661 gagaaaggct agaaaaagca agatctggga tatttcctta gaatccttgc ctgaaggcca
      721 agaagatgag gaagttaatt ttctcatcca gagtggtggt atcgagctag aaaattgtga
      781 agatctttct agttctaaaa catctgttac ctccttacct tggggtttca agactgggtg
      841 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggc
      901 gtggccacca tggctcctaa gaagaagagg aaggtgatga gccagttcga catcctgtgc
      961 aagacccccc ccaaggtgct ggtgcggcag ttcgtggaga gattcgagag gcccagcggc
     1021 gagaagatcg ccagctgtgc cgccgagctg acctacctgt gctggatgat cacccacaac
     1081 ggcaccgcca tcaagagggc caccttcatg agctacaaca ccatcatcag caacagcctg
     1141 agcttcgaca tcgtgaacaa gagcctgcag ttcaagtaca agacccagaa ggccaccatc
     1201 ctggaggcca gcctgaagaa gctgatcccc gcctgggagt tcaccatcat cccttacaac
     1261 ggccagaagc accagagcga catcaccgac atcgtgtcca gcctgcagct gcagttcgag
     1321 agcagcgagg aggccgacaa gggcaacagc cacagcaaga agatgctgaa ggccctgctg
     1381 tccgagggcg agagcatctg ggagatcacc gagaagatcc tgaacagctt cgagtacacc
     1441 agcaggttca ccaagaccaa gaccctgtac cagttcctgt tcctggccac attcatcaac
     1501 tgcggcaggt tcagcgacat caagaacgtg gaccccaaga gcttcaagct ggtgcagaac
     1561 aagtacctgg gcgtgatcat tcagtgcctg gtgaccgaga ccaagacaag cgtgtccagg
     1621 cacatctact ttttcagcgc cagaggcagg atcgaccccc tggtgtacct ggacgagttc
     1681 ctgaggaaca gcgagcccgt gctgaagaga gtgaacagga ccggcaacag cagcagcaac
     1741 aagcaggagt accagctgct gaaggacaac ctggtgcgca gctacaacaa ggccctgaag
     1801 aagaacgccc cctaccccat cttcgctatc aagaacggcc ctaagagcca catcggcagg
     1861 cacctgatga ccagctttct gagcatgaag ggcctgaccg agctgacaaa cgtggtgggc
     1921 aactggagcg acaagagggc ctccgccgtg gccaggacca cctacaccca ccagatcacc
     1981 gccatccccg accactactt cgccctggtg tccaggtact acgcctacga ccccatcagc
     2041 aaggagatga tcgccctgaa ggacgagacc aaccccatcg aggagtggca gcacatcgag
     2101 cagctgaagg gcagcgccga gggcagcatc agataccccg cctggaacgg catcatcagc
     2161 caggaggtgc tggactacct gagcagctac atcaacaggc ggatctgaga attcgatatc
     2221 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2401 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2461 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2521 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2581 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2641 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2701 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2761 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2821 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     2881 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     2941 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3001 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3061 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3121 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3181 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3241 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3301 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3361 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3421 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3481 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3541 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3661 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3781 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3901 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3961 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4021 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4261 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
