LOCUS       sequence-462399-        5371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0050m_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 5371)
  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 5371)
  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: AiP13776 - pAAV-AiE0050m_3xC2-minBG-iCre-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1062..1082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2198..2218)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2405..2516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2634..2774
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2634..2763
                     /label=AAV2 ITR
     rep_origin      2849..3304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2936..2955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3146..3167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3321..3340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3440..3462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3500..3518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3586..3690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3691..3759,3760..4551)
                     /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(3909..3928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4722..5310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5211..5230
                     /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 gcgtaggcaa cttgttaaaa atctcagtca aacttacata
      181 ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag tttcttattc
      241 agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa agtcacaaat
      301 acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact tttattttgc
      361 ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca gcaggaaata
      421 ggcaacttgt taaaaatctc agtcaaactt acatactata tagaacagca tggtgaattt
      481 agggcacatg gatataaaat ggaagtttct tattcagtag cagcaacttg tgggcacagg
      541 agttggcaaa gataaaaatg tccaaagtca caaatacaat gtatagttag tcataggtgc
      601 tgttatttgc ctcaaaaaat agacttttat tttgcctttc ttttctttaa ccacactcaa
      661 aattagagaa cagagacaaa acccagcagg aaataggcaa cttgttaaaa atctcagtca
      721 aacttacata ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag
      781 tttcttattc agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa
      841 agtcacaaat acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact
      901 tttattttgc ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca
      961 gcaggaaatg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1021 tgcttctggg atccagatct ttcgaagcta gcaccatggt gcccaagaag aagaggaaag
     1081 tctccaacct gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg
     1141 atgaagtcag gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca
     1201 cctggaagat gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca
     1261 ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca
     1321 gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga
     1381 gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca
     1441 gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg
     1501 actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc
     1561 tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca
     1621 gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca
     1681 agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg
     1741 tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc
     1801 gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc
     1861 tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc
     1921 agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca
     1981 gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg
     2041 tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg
     2101 atggggacta atgaggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     2161 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2221 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2281 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2341 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2401 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2461 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2521 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2581 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2641 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2701 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2761 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2821 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2881 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2941 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3001 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3061 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3121 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3181 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3241 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3301 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3481 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4741 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4801 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4861 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4921 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4981 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5041 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5101 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5161 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5221 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5281 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5341 ctggcctttt gctggccttt tgctcacatg t
//
