LOCUS       sequence-461933-        5166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0526h_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 5166)
  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 5166)
  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: AiP15127 -
            pAAV-AiE0526h_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..5166
                     /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      1102..1111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1114..1134
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2200..2311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2429..2569
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2429..2558
                     /label=AAV2 ITR
     rep_origin      2644..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2941..2962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3116..3135)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3235..3257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3295..3313)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3381..3485
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3486..3554,3555..4346)
                     /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(3704..3723)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4517..5105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5006..5025
                     /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 gcgtactaag atgtttatgg caatcgccct gcaagtgtct
      181 atttaattcc tgaaaatgtc atttactggt gtgtttctgt aaaaatcatt tctgaaaagt
      241 tttgtcagct tggcttagca gctgggggaa aaaagcagca aaatgagtcc cctcaatcaa
      301 gcctgtgatt ttttattttt tcagactcat gaatcctcct gagaattagg cttatgacaa
      361 gcacccaccc cctccaccta ctccctggtt taaaccaggt gcctgctgac acgaaggaag
      421 attccggggc attttgacta agatgtttat ggcaatcgcc ctgcaagtgt ctatttaatt
      481 cctgaaaatg tcatttactg gtgtgtttct gtaaaaatca tttctgaaaa gttttgtcag
      541 cttggcttag cagctggggg aaaaaagcag caaaatgagt cccctcaatc aagcctgtga
      601 ttttttattt tttcagactc atgaatcctc ctgagaatta ggcttatgac aagcacccac
      661 cccctccacc tactccctgg tttaaaccag gtgcctgctg acacgaagga agattccggg
      721 gcattttgac taagatgttt atggcaatcg ccctgcaagt gtctatttaa ttcctgaaaa
      781 tgtcatttac tggtgtgttt ctgtaaaaat catttctgaa aagttttgtc agcttggctt
      841 agcagctggg ggaaaaaagc agcaaaatga gtcccctcaa tcaagcctgt gattttttat
      901 tttttcagac tcatgaatcc tcctgagaat taggcttatg acaagcaccc accccctcca
      961 cctactccct ggtttaaacc aggtgcctgc tgacacgaag gaagattccg gggcattttg
     1021 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
     1081 gatccagatc tttcgaagct agccaccatg gtgcccaaga agaagaggaa agtctccaac
     1141 ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg atgccacctc tgatgaagtc
     1201 aggaagaacc tgatggacat gttcagggac aggcaggcct tctctgaaca cacctggaag
     1261 atgctcctgt ctgtgtgcag atcctgggct gcctggtgca agctgaacaa caggaaatgg
     1321 ttccctgctg aacctgagga tgtgagggac tacctcctgt acctgcaagc cagaggcctg
     1381 gctgtgaaga ccatccaaca gcacctgggc cagctcaaca tgctgcacag gagatctggc
     1441 ctgcctcgcc cttctgactc caatgctgtg tccctggtga tgaggagaat cagaaaggag
     1501 aatgtggatg ctggggagag agccaagcag gccctggcct ttgaacgcac tgactttgac
     1561 caagtcagat ccctgatgga gaactctgac agatgccagg acatcaggaa cctggccttc
     1621 ctgggcattg cctacaacac cctgctgcgc attgccgaaa ttgccagaat cagagtgaag
     1681 gacatctccc gcaccgatgg tgggagaatg ctgatccaca ttggcaggac caagaccctg
     1741 gtgtccacag ctggtgtgga gaaggccctg tccctggggg ttaccaagct ggtggagaga
     1801 tggatctctg tgtctggtgt ggctgatgac cccaacaact acctgttctg ccgggtcaga
     1861 aagaatggtg tggctgcccc ttctgccacc tcccaactgt ccacccgggc cctggaaggg
     1921 atctttgagg ccacccaccg cctgatctat ggtgccaagg atgactctgg gcagagatac
     1981 ctggcctggt ctggccactc tgccagagtg ggtgctgcca ccgacatggc cagggctggt
     2041 gtgtccatcc ctgaaatcat gcaggctggt ggctggacca atgtgaacat tgtgatgaac
     2101 tacatcagaa acctggactc tgagactggg gccatggtga ggctgctcga agatggggac
     2161 tgaggcgcgc cgaattcaag cttctcgaga gatcttcgac tgtgccttct agttgccagc
     2221 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2281 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2341 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2401 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2461 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2521 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2581 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2641 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2701 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2821 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2941 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     3001 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     3061 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3121 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3181 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3241 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3301 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3361 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3481 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3541 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3601 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3661 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3721 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3781 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3841 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3901 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3961 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4021 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4081 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4141 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4201 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4261 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4321 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4381 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4441 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5161 acatgt
//
