LOCUS       sequence-433042-        5432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2404m 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 5432)
  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 5432)
  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: AiP1712 - pAAV-AiE2404m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(148..167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     267..289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(327..345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        413..517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(518..586,587..1378)
                     /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(736..755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1549..2137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2038..2057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2199..2328
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3642..3651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3648..3650,3651..3653,3654..4364)
                     /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(3693..3714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3954..3973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4301..4322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4457..4477)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4664..4775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4893..5033
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4893..5022
                     /label=AAV2 ITR
     rep_origin      join(5108..5432,1..131)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5195..5214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5405..5426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
       61 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
      121 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
      241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
      301 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      361 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
      421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2161 acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg
     2221 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     2281 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc
     2341 gttgtaagag taataatcgt cctgtattat caagagttga aatgtgtcta ctgggaccag
     2401 tcttgatgcc gatattttct gtcttgccca atgtgcaaac agggcaagcc tgaactcgta
     2461 attccatgcc ataaaacctg gtttaatttc actgacattc caagtgtaac tcaagtgcct
     2521 ttctacaaag acttactcat atctcatgaa gttagtgtat ttcattagca tggaaatcca
     2581 aaatgacctg tttgatgtcc agccattcag taagtaggat acaggaaaga tctgttaaag
     2641 ataaaattgt caaaatggag agaatcacat aatttatttt agctttcgtt gtaaacaacc
     2701 accttttttt ttttttgtac tatcaaactc atgatttcct gacagctgaa gacatctatg
     2761 cccatggcct gcttactttt taaatttcta ccttgcttga cttctttctc tattcaaaag
     2821 aaaacactaa gcaaggggga aaaagaaagt atgtccacaa gcagacagag actgggatga
     2881 agcctgagct cagcatatac gtttcaactg ctttttaagg cactgggttt tgttttcaat
     2941 gtacaggtgt ttatctttat ttagaactaa gtttaaatgt gacacagatt cattgcaatt
     3001 tcccactgtc ctgtatctga aaagcttcca agttccctga cgctcttctt tgtccctggg
     3061 tatcatgatt tcagtgtttt tcaaatctat tcctcagctc cagtgtttaa tgtgtgtaca
     3121 gggatttaac cataacgtat gtaaagaaaa agctagtact tgtctattaa aaaaaaatag
     3181 ctacttcttt ctattggttt ctatgtactg tgacaaattt aagcccgatt tttttcaacc
     3241 tatgggtaat gtcataggta taagatagtg ggtacagcat gaaataataa gaagaaatta
     3301 ttagtaaaca ttcattatga tgcattgtat gccaagtact taatggataa tttgtataca
     3361 tatcctcaag taatctgaat aattaatctc aaagatggaa agtttcattt tatagactaa
     3421 agagatgcac agaggccatg ccagttcctt gagtgtccct taggaggaca agcagataca
     3481 aatccaagca ttgggctaga gaaacactgc aacagtgaag agcattggta gctcttccag
     3541 aggacacgga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     3601 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
     3661 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3721 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3781 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3841 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     3901 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3961 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4021 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4081 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     4141 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     4201 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     4261 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     4321 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     4381 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     4441 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4501 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4561 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4621 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     4681 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4741 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4801 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4861 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     4921 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4981 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     5041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     5101 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5161 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     5221 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5281 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     5341 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5401 aatagtggac tcttgttcca aactggaaca ac
//
