LOCUS       sequence-474119-        4985 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 4985)
  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 4985)
  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: AiP14587 - pAAV-AiE0330h-minBG-iCre(R297T)-BGH...
FEATURES             Location/Qualifiers
     source          1..4985
                     /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      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             933..953
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2019..2130
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2248..2388
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2248..2377
                     /label=AAV2 ITR
     rep_origin      2463..2918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2550..2569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2760..2781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2935..2954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3054..3076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3114..3132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3200..3304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3305..3373,3374..4165)
                     /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(3523..3542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4336..4924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4825..4844
                     /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 tgcttctggg
      901 atccagatct ttcgaagcta gccaccatgg tgcccaagaa gaagaggaaa gtctccaacc
      961 tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct gatgaagtca
     1021 ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac acctggaaga
     1081 tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac aggaaatggt
     1141 tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc agaggcctgg
     1201 ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg agatctggcc
     1261 tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc agaaaggaga
     1321 atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact gactttgacc
     1381 aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac ctggccttcc
     1441 tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc agagtgaagg
     1501 acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc aagaccctgg
     1561 tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg gtggagagat
     1621 ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc cgggtcagaa
     1681 agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc ctggaaggga
     1741 tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg cagagatacc
     1801 tggcctggtc tggccactct gccagagtgg gtgctgccac cgacatggcc agggctggtg
     1861 tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt gtgatgaact
     1921 acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgaa gatggggact
     1981 gaggcgcgcc gaattcaagc ttctcgagag atcttcgact gtgccttcta gttgccagcc
     2041 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2101 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2161 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2221 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2281 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2341 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2401 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2461 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2521 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2641 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2761 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2821 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2881 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2941 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3001 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3061 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3121 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3181 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3241 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3301 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3361 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3421 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3481 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3541 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3601 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3661 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3721 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3781 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3841 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3901 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3961 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4021 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4081 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4141 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4201 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4261 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4321 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4381 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4441 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4501 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4561 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4621 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4681 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4741 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4801 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4861 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4921 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4981 catgt
//
