LOCUS       sequence-466304-        5092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2442m 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 5092)
  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 5092)
  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: AiP1771 - pAAV-AiE2442m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      440..449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(446..448,449..451,452..1162)
                     /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(491..512)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(752..771)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1099..1120
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1255..1275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1462..1573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1691..1831
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1691..1820
                     /label=AAV2 ITR
     rep_origin      1906..2361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1993..2012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2203..2224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2378..2397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2497..2519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2557..2575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2643..2747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2748..2816,2817..3608)
                     /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(2966..2985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3779..4367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4268..4287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4429..4558
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gggttgggct agctgtcaga agcacactct tttttgttgt gtgtgatatg aaggttatgg
       61 gaaggtgtgt gtgtgtgtgt gtgcacgcgc gcgcgcgtgc gtgtgcatgt gtttgcatgc
      121 acacatgtgg gtatatgtgc acaggccaga tgccaacctc gagtttcttc ctcatttgtt
      181 ttctgccttt attttttcag acaggatctt ttactaagcc tagagctctt tgattgtgct
      241 aggctgcctg gctagtgagc ggcagagacc cttttgcctc tgtctctaca gtactgagat
      301 ttcaggcatg tgccaccttg gctagctctt ctcagtgaga taggcggagc tcgggctggg
      361 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc
      421 gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      481 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      541 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
      601 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
      661 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      721 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      781 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      841 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      901 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
      961 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1021 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1081 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1141 cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca
     1201 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1261 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1321 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1381 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1441 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1501 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1561 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1621 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     1681 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     1741 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     1801 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     1861 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     1921 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1981 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2041 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2101 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2161 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2221 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2281 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2341 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2401 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2461 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2521 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     2581 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     2641 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2701 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2761 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2821 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2881 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2941 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3001 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3061 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3121 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3181 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3241 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3301 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3361 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3421 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3481 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3541 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3601 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3661 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3721 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3781 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3841 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3901 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3961 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4021 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4081 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4141 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4201 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4261 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4321 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4381 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     4441 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     4501 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     4561 ggccgcacgc gttgtaagag taataatcgg aagctctttt tctttcacat ctgctgagtc
     4621 atctctgtac gtgactgtat gtctctctac acctagagag acttctctgc ccaacccatt
     4681 tcaccatagt cccagaatta tctttatcat ctgggcccca acttgtcagt ggctatttat
     4741 gttgaagagc caggcatagt gatccatatc tataatctca gcattcagga agctgaagca
     4801 ggataattgc aagtttgaag tcaacctggc aaatttaagg ccagccttaa atacatactg
     4861 catagtgaat tcgggaccca gccctagtgg tggagaggca tcatggtgac atacaaattt
     4921 gggtgtattc ttccaaataa agaaagctgc tgagaatgat gcttcagttc tgttgtgttt
     4981 gggctccttg agcatctctc agaagggagg gaagagtgac tcacttcatt ggcaacttca
     5041 ggaagggttc tggcagcaca cagacacttg aggaccttcc cgcatctctc tc
//
