LOCUS       sequence-432547-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2362m 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 4922)
  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 4922)
  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: AiP1680 - pAAV-AiE2362m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(484..503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     603..625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(663..681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        749..853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(854..922,923..1714)
                     /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(1072..1091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1885..2473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2374..2393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2535..2664
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3468..3477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3474..3476,3477..3479,3480..4190)
                     /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(3519..3540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3780..3799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4127..4148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4283..4303)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4490..4601
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4719..4859
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4719..4848
                     /label=AAV2 ITR
ORIGIN
        1 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
      361 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
      481 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
      661 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      841 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      901 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      961 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1021 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1081 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1141 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1201 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1261 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1321 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1381 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1441 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1501 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1561 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1621 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1681 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1741 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1801 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1861 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2521 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     2581 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     2641 ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat aatcgtgatc
     2701 cccctgacgg ctgtagagag gcaggctttg tttttacttg tatgcagtcg ttttagccag
     2761 tctctcgtca tggctgcata ctcctgccag ctgtacccag agaacaaaga gtacctcatg
     2821 attacagcca aaaccggatg gaagcattta cagcagctgt tcgacatggg ccagaaagcc
     2881 gttgaagaaa tctggtttga aactgccaag tcctatgaat ctgagatatc catgtgactg
     2941 ccgcggtaaa aaaattaaat cgctcaaaac tccccggtgg tgcacatgcg ctcggcgcta
     3001 tgcacccaca atgttctgga ttcccgagtg aaagacacac acgcacaacc ttatttttaa
     3061 tatgccctaa gcaggtcaat tgccgggcca ctccctaact ccacttggtt aacgcacact
     3121 ccacttcaat attcctgagt tattacttac taaaatctat atattatctt tgtgccctgg
     3181 acctggctgg gcagccctcc tgggcagctt tccccctgca cctacatgtc ggcggtcttt
     3241 cccttctgta tctttctggc gtggtcctat gcccctcttg tggcaaaatc ttctcttcct
     3301 actctctctg tctcttccca ggaatcctaa aagtcccacc tctgtctctg cccagtcatt
     3361 ggccgctggt aacggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt
     3421 acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga
     3481 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3541 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3601 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3661 ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     3721 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3781 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3841 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3901 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     3961 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     4021 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4081 gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4141 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg
     4201 gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa
     4261 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4321 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4381 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4441 gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct
     4501 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4561 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4621 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4681 agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4741 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4801 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4861 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4921 aa
//
