LOCUS       sequence-474096-        5864 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 5864)
  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 5864)
  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: AiP12784 - pAAV-AiE0330h-minBG-iCre-WPRE-HGHpA...
FEATURES             Location/Qualifiers
     source          1..5864
                     /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      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             917..937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1991..2579
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2044..2064)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2462..2473)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2611..3087
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2810..2829)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3127..3267
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3127..3256
                     /label=AAV2 ITR
     rep_origin      3342..3797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3429..3448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3639..3660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3814..3833)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3933..3955
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3993..4011)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4079..4183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4184..4252,4253..5044)
                     /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(4402..4421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5215..5803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5704..5723
                     /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 atccgccacc atggtgccca agaagaagag gaaagtctcc aacctgctga ctgtgcacca
      961 aaacctgcct gccctccctg tggatgccac ctctgatgaa gtcaggaaga acctgatgga
     1021 catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc tgtctgtgtg
     1081 cagatcctgg gctgcctggt gcaagctgaa caacaggaaa tggttccctg ctgaacctga
     1141 ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga agaccatcca
     1201 acagcacctg ggccagctca acatgctgca caggagatct ggcctgcctc gcccttctga
     1261 ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg atgctgggga
     1321 gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca gatccctgat
     1381 ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca ttgcctacaa
     1441 caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct cccgcaccga
     1501 tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca cagctggtgt
     1561 ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct ctgtgtctgg
     1621 tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg gtgtggctgc
     1681 cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg aggccaccca
     1741 ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct ggtctggcca
     1801 ctctgccaga gtgggtgctg ccagggacat ggccagggct ggtgtgtcca tccctgaaat
     1861 catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca gaaacctgga
     1921 ctctgagact ggggccatgg tgaggctgct cgaggatggg gactaagaat tcgatatcaa
     1981 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2041 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2101 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2161 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2221 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2281 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2341 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2401 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2461 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2521 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2581 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     2641 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     2701 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     2761 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     2821 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     2881 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     2941 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3001 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3061 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3121 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3181 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3241 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3301 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3541 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3661 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3721 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3781 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3841 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3901 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3961 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4021 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4381 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4561 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5041 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5341 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5821 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
