LOCUS       sequence-432561-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2468m 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 4851)
  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 4851)
  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: AiP1777 - pAAV-AiE2468m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   274..403
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1136..1145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1142..1144,1145..1147,1148..1858)
                     /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(1187..1208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1448..1467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1795..1816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1951..1971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2158..2269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2387..2527
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2387..2516
                     /label=AAV2 ITR
     rep_origin      2602..3057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2689..2708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2899..2920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3074..3093)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3193..3215
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3253..3271)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3339..3443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3444..3512,3513..4304)
                     /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(3662..3681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4475..4851,1..212)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
       61 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      241 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
      301 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
      361 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgttgt
      421 aagagtaata atcgagagaa aaggaaacgc tagaaaagct gagctgcaag cccaggctgc
      481 catttgaaag agcagttgag gaagatctga atgaaaaact ctgcaaattt atgctaattg
      541 tgttcagttc catgagtttc atcccattga gtgggagcca ggccacctga ttgcttctcc
      601 ttttggtaaa gaacatctca gatgaaccag ctgtcatgga catggttcag aaaattggcc
      661 ctaaaaagaa acctgggcca tctgtgtgtg ctgttctatt cataggatgc cctccaaatc
      721 acatatttct attaggaacc tcacagcaag ggcctctacc atgggagcaa tagagaacat
      781 tggtcatcat catctattgc actcccttgg ggtgggagtg ctgtgatttc tagcaaagag
      841 aactcagggc tgaggtagat aacaaagaca acttggaatg tctaacgtga ggaagcatcc
      901 ttctcttcct tttgctttgt cagattctgt aactgacaag aaattggatg tgagattgtg
      961 aaagtatttt gtgtgcattg aacaagaacg tttgcaaggt tccagggtga gcctcgaagc
     1021 ccgggcctac cctgcctggg cggagctcgg gctgggcata aaagtcaggg cagagccatc
     1081 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
     1141 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1201 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1261 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1321 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1441 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1561 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1621 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1681 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1741 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1801 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1861 agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat
     1921 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1981 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2041 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2101 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
     2161 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2221 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2281 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2341 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
     2401 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2461 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2521 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2581 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2641 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2701 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2761 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2821 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2881 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     2941 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3001 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3061 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3121 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3181 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3241 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3301 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3361 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3421 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3481 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3541 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3601 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3661 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3721 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3781 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3841 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3901 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3961 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4021 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4081 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4141 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4201 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4261 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4321 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4381 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4441 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4501 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4561 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4621 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4681 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4741 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4801 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga t
//
