LOCUS       sequence-470785-        4974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0170h_3xC2 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 4974)
  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 4974)
  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: AiP15285 -
            pAAV-AiE0170h_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             922..942
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2008..2119
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2237..2377
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2237..2366
                     /label=AAV2 ITR
     rep_origin      2452..2907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2539..2558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2749..2770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2924..2943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3043..3065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3103..3121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3189..3293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3294..3362,3363..4154)
                     /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(3512..3531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4325..4913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4814..4833
                     /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 gcgtctgtct tttagctaat ctcacaatat tcaatatttc
      181 ttgccaggat tattgcaatt ccctcctgtt aagtctcccc acttaaactc cagctgcacc
      241 aatctacact ttacaattct gcagagatgt ctttttaaca tgcaaatatg cttatattat
      301 ttctttgctt acacattcca ataactcctc taaataaaac tcttaagttg cttagactga
      361 cacaaggctc tcctgtcttt tagctaatct cacaatattc aatatttctt gccaggatta
      421 ttgcaattcc ctcctgttaa gtctccccac ttaaactcca gctgcaccaa tctacacttt
      481 acaattctgc agagatgtct ttttaacatg caaatatgct tatattattt ctttgcttac
      541 acattccaat aactcctcta aataaaactc ttaagttgct tagactgaca caaggctctc
      601 ctgtctttta gctaatctca caatattcaa tatttcttgc caggattatt gcaattccct
      661 cctgttaagt ctccccactt aaactccagc tgcaccaatc tacactttac aattctgcag
      721 agatgtcttt ttaacatgca aatatgctta tattatttct ttgcttacac attccaataa
      781 ctcctctaaa taaaactctt aagttgctta gactgacaca aggctctcga gctcgggctg
      841 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
      901 tcgaagctag ccaccatggt gcccaagaag aagaggaaag tctccaacct gctgactgtg
      961 caccaaaacc tgcctgccct ccctgtggat gccacctctg atgaagtcag gaagaacctg
     1021 atggacatgt tcagggacag gcaggccttc tctgaacaca cctggaagat gctcctgtct
     1081 gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt ccctgctgaa
     1141 cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc tgtgaagacc
     1201 atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct gcctcgccct
     1261 tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa tgtggatgct
     1321 ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca agtcagatcc
     1381 ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct gggcattgcc
     1441 tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga catctcccgc
     1501 accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt gtccacagct
     1561 ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg gatctctgtg
     1621 tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa gaatggtgtg
     1681 gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat ctttgaggcc
     1741 acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct ggcctggtct
     1801 ggccactctg ccagagtggg tgctgccacc gacatggcca gggctggtgt gtccatccct
     1861 gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta catcagaaac
     1921 ctggactctg agactggggc catggtgagg ctgctcgaag atggggactg aggcgcgccg
     1981 aattcaagct tctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2161 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2221 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2281 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2341 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2401 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2461 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2521 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2581 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2641 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2701 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2761 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2821 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2881 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2941 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3001 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3061 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3121 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3181 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3241 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3301 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3361 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3421 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3481 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3541 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3601 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3661 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3721 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3781 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3841 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3901 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3961 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4021 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4081 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4141 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4201 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4261 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4321 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4381 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4441 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4501 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4561 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4621 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4681 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4741 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4801 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4861 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4921 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
