LOCUS       sequence-471054-        4971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0464m_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 4971)
  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 4971)
  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: AiP20138 -
            pAAV-AiE0464m_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..4971
                     /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      907..916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             919..939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2005..2116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2234..2374
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2234..2363
                     /label=AAV2 ITR
     rep_origin      2449..2904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2536..2555)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2746..2767
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2921..2940)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3040..3062
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3100..3118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3186..3290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3291..3359,3360..4151)
                     /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(3509..3528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4322..4910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4811..4830
                     /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 gcgttttgtg tctgctattg catttacata gtgacatagg
      181 cttacaggat attgtaaatg tcatccctag atacatttat gtaacctttt tttaaaataa
      241 aactctaaca aatgttagtt attgtgcact atggttttga atatttgaat ttgctttgct
      301 cagatctaaa taacttctga taagtttggc attcaggaaa gagtaaatgt aaatgtagaa
      361 atgcttggaa ttttgtgtct gctattgcat ttacatagtg acataggctt acaggatatt
      421 gtaaatgtca tccctagata catttatgta accttttttt aaaataaaac tctaacaaat
      481 gttagttatt gtgcactatg gttttgaata tttgaatttg ctttgctcag atctaaataa
      541 cttctgataa gtttggcatt caggaaagag taaatgtaaa tgtagaaatg cttggaattt
      601 tgtgtctgct attgcattta catagtgaca taggcttaca ggatattgta aatgtcatcc
      661 ctagatacat ttatgtaacc tttttttaaa ataaaactct aacaaatgtt agttattgtg
      721 cactatggtt ttgaatattt gaatttgctt tgctcagatc taaataactt ctgataagtt
      781 tggcattcag gaaagagtaa atgtaaatgt agaaatgctt ggaatgagct cgggctgggc
      841 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      901 aagctagcca ccatggtgcc caagaagaag aggaaagtct ccaacctgct gactgtgcac
      961 caaaacctgc ctgccctccc tgtggatgcc acctctgatg aagtcaggaa gaacctgatg
     1021 gacatgttca gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg
     1081 tgcagatcct gggctgcctg gtgcaagctg aacaacagga aatggttccc tgctgaacct
     1141 gaggatgtga gggactacct cctgtacctg caagccagag gcctggctgt gaagaccatc
     1201 caacagcacc tgggccagct caacatgctg cacaggagat ctggcctgcc tcgcccttct
     1261 gactccaatg ctgtgtccct ggtgatgagg agaatcagaa aggagaatgt ggatgctggg
     1321 gagagagcca agcaggccct ggcctttgaa cgcactgact ttgaccaagt cagatccctg
     1381 atggagaact ctgacagatg ccaggacatc aggaacctgg ccttcctggg cattgcctac
     1441 aacaccctgc tgcgcattgc cgaaattgcc agaatcagag tgaaggacat ctcccgcacc
     1501 gatggtggga gaatgctgat ccacattggc aggaccaaga ccctggtgtc cacagctggt
     1561 gtggagaagg ccctgtccct gggggttacc aagctggtgg agagatggat ctctgtgtct
     1621 ggtgtggctg atgaccccaa caactacctg ttctgccggg tcagaaagaa tggtgtggct
     1681 gccccttctg ccacctccca actgtccacc cgggccctgg aagggatctt tgaggccacc
     1741 caccgcctga tctatggtgc caaggatgac tctgggcaga gatacctggc ctggtctggc
     1801 cactctgcca gagtgggtgc tgccaccgac atggccaggg ctggtgtgtc catccctgaa
     1861 atcatgcagg ctggtggctg gaccaatgtg aacattgtga tgaactacat cagaaacctg
     1921 gactctgaga ctggggccat ggtgaggctg ctcgaagatg gggactgagg cgcgccgaat
     1981 tcaagcttct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     2221 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2281 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2341 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2401 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2461 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2521 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2581 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2641 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2701 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2761 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2821 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2881 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2941 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3001 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3061 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3121 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3181 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3301 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3361 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3421 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3481 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3541 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3601 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3661 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3721 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3781 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3841 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3901 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3961 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4021 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4081 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4141 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4201 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4261 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4441 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4681 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
