LOCUS       sequence-492625-        5047 bp    DNA     circular SYN 12-NOV-2025
DEFINITION  AiE2575m 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 5047)
  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 5047)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 12, 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: AiP20168 - pAAV-AiE2575m-minBG-iCre-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1395..1404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1407..1427
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2524..2544)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2731..2842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2960..3100
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2960..3089
                     /label=AAV2 ITR
     rep_origin      3175..3630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3262..3281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3472..3493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3647..3666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3766..3788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3826..3844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4017..4085,4086..4877)
                     /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(4235..4254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcat tactgcagtc acaccgaagg gaagattcaa cgagaatggg aaattatgag
      841 aaatgcccac acttcctatt tcctatatct attttagttg gtggcaatta caatttacta
      901 cgcaacactc aaaatataaa cggcactatg tgaaaggata taacagtgac agatttgcag
      961 cctcaaagcg tgacttcaaa atattggtgc cgaaaaggtt tcctgttccg gaaggcagtt
     1021 gagactacaa agcccacgca ggtgaaggcg ctggcaggag cgagtgctct gcgtaccctg
     1081 gatgttgagc aaagctgtgc agggtagatg ggaggcgccg cccggcccca tatggagccg
     1141 accttccgtg ccaatcgcac tatgctatgg cacacacagc gagctgagcg aggaaatcct
     1201 gcgaagcagg gaaactgaga gcataaggcc cggagttgaa aatcaagaag gaaagccaag
     1261 agtttgaggg aaatgcagag gcagggttat gtgaggagaa catgccagtg tcctgtctct
     1321 ttagccccgg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1381 tgcttctggg atccgccacc atggtgccca agaagaagag gaaagtctcc aacctgctga
     1441 ctgtgcacca aaacctgcct gccctccctg tggatgccac ctctgatgaa gtcaggaaga
     1501 acctgatgga catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc
     1561 tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa caacaggaaa tggttccctg
     1621 ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga
     1681 agaccatcca acagcacctg ggccagctca acatgctgca caggagatct ggcctgcctc
     1741 gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg
     1801 atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca
     1861 gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca
     1921 ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct
     1981 cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca
     2041 cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct
     2101 ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg
     2161 gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg
     2221 aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct
     2281 ggtctggcca ctctgccaga gtgggtgctg ccagggacat ggccagggct ggtgtgtcca
     2341 tccctgaaat catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca
     2401 gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaggatggg gactaagaat
     2461 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2521 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2581 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2641 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2701 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     2761 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2821 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2881 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     2941 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3001 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3061 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3121 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3481 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3541 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3601 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttc
//
