LOCUS       sequence-432590-        4987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2446m 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 4987)
  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 4987)
  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: AiP1933 - pAAV-AiE2446m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   548..677
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(739..1327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(819..838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1498..2289,2290..2358))
                     /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     2121..2140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2359..2463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2531..2549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2587..2609)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2709..2728
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2745..3200)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2882..2903)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3094..3113
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3275..3415
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3286..3415
                     /label=AAV2 ITR
     polyA_signal    3533..3644
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3831..3851
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3944..4654,4655..4657,4658..4660))
                     /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(3986..4007)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4335..4354
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4594..4615
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4657..4666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tctcatgctc atggattggc agaaccaaca tagtaaaagt ggccatcctg ccaagggcaa
       61 tctacagatt cactgcagta cccatcaaaa tcccaacaca gttcttcaga gacatggaaa
      121 gagcaattct caaattcatc tggaaaggca aaaagcccag aatagctaaa aacagttctt
      181 aacaatgcca agaatttggc atttactcat gagatctggt acaactggat tgaaatttga
      241 ggaaaaagta ggttaccacc tgtcatttaa gcagtgctgc attttctata tttaggtttt
      301 aatatgtttc attttgtttt atcatttatt tttagcacat ttgatgctcc cagttctccc
      361 ctcgttcttt ctttgaatct taggtcttca atttagaaat tgccattaat gtactgtttt
      421 ataaatcgac tatgatgcat cctgttgaaa cctactgcca tcttttcttt accaaggatt
      481 tgaatactct agtcattttt atttttcatt ttcaaaattt ttattattga ttattttttg
      541 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
      601 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
      661 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1141 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1381 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1441 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1501 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1561 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1621 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1681 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1741 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1801 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1861 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1921 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1981 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2041 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2101 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2161 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2221 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2281 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2341 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2461 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2521 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2581 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2641 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2701 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     2761 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     2821 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     2881 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2941 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     3001 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3061 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3121 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3181 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     3241 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
     3301 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     3361 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     3421 cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct
     3481 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
     3541 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
     3601 gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag
     3661 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     3721 cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag
     3781 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     3841 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     3901 gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca
     3961 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     4021 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     4081 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     4141 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     4201 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     4261 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     4321 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     4381 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     4441 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     4501 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     4561 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     4621 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     4681 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     4741 ttatgcccag cccgagctcc ggcaaagtgg ctgcatataa aattaactca attaaatcag
     4801 tgactttcct ttataaaaat gataaacagg ttgagaaaga aattagggaa aaaatccctt
     4861 cataatagca acaaataata taagatatct taagggtaac tctaaccaaa caactgaaag
     4921 atttgtatgt caataacttc cagcctcgca agaatgaaat cgaagatctc agtaaaatga
     4981 agagatc
//
