LOCUS       sequence-432553-        5363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2405m 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 5363)
  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 5363)
  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: AiP1713 - pAAV-AiE2405m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     292..313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(467..486)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     586..608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(646..664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        732..836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(837..905,906..1697)
                     /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(1055..1074)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1868..2456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2357..2376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2518..2647
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3892..3901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3898..3900,3901..3903,3904..4614)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3943..3964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4204..4223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4551..4572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4707..4727)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4914..5025
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5143..5283
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5143..5272
                     /label=AAV2 ITR
     rep_origin      join(5358..5363,1..450)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
       61 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      181 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
      241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      301 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
      361 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      421 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
      481 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      541 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      601 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
      661 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      721 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      781 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      841 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      901 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      961 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1021 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1081 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1141 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1201 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1261 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1321 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1381 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1441 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1501 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1561 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1621 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1681 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1741 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1801 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1861 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1921 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1981 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2041 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2101 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2161 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2221 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2281 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2341 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2401 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2461 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
     2521 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
     2581 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     2641 ggttcctgcg gccgcacgcg ttgtaagagt aataatcgtg tcttctacct acctctgcaa
     2701 cgtttaatga tcaatagtcc agattccaca ctcacttttt aggaggccct attccgacag
     2761 ggacaaacga gacactgata gggaaaatga ctgtgacagt tgacacctga aatgcactga
     2821 gggaaagaca gtatgtgatt tagagaagat gagaatgtca tagttggagg gaaataagat
     2881 gatgtagaac caggatttgg aataaataat gaaaatatca aagacaaaac aaagatgcag
     2941 aaactgggca gtaaaacagt ggtcttcagc ttggagggga acattattcg tgggaagtcc
     3001 tcaatggtac ctcgttcaaa agaatttcca caaaagacat tcaccgattt caaagtgtgt
     3061 ttgttatgta ataagaacat tctttatctt cccaggcatt tgttgaaata gactccgtgc
     3121 acaactctta tgtgaaatgc atttgaacag aaaaaataaa atggactgtg ttctacatct
     3181 gctttggaaa aggctggtga ggaagaaagc tcagagctca gtaggcacag caagcagagg
     3241 ctcggggacc cacacaacca cagagaggcc actgtgccat gaatggggaa gacgggatgt
     3301 gttggtttat ataaatatca aaaaaaattt atatacccca cttagacttg atcaactgac
     3361 ttgtctcttt aagcccacct agttggtggg tagaagaaat gtctagaacg ttattgtcat
     3421 catccctttc aatggcattt gacaaatggg tgcttctggg agagggcagc tgaactattt
     3481 cccttccatt caccgttgct tttgatgtca ccatctgcct tttcatgtat ttttattcct
     3541 aaggaggaat aagtaagtga ttcttagtca cagagttagg atctgtggtt tagaaaaatg
     3601 ctgagtccag tcaaatgcga ttacgtttgc aaacaaaggt gccactaatt gtgtgcaaga
     3661 cagtcccttg acaaggggac atctttcata tcatatccac tgcctccaca tagccataag
     3721 cctctagaca ctgcaagaat ttctgaccac caagcttgct tatcagtgaa acttttgtta
     3781 taccaatgga aagatacgga gctcgggctg ggcataaaag tcagggcaga gccatctatt
     3841 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
     3901 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4021 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4081 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     4141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4321 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     4381 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     4441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4501 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4561 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc
     4621 gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     4681 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4741 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4801 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4861 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     4921 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4981 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5041 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5101 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     5161 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     5221 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     5281 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     5341 gtcaaagcaa ccatagtacg cgc
//
