LOCUS       sequence-466300-        5006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2419m 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 5006)
  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 5006)
  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: AiP1751 - pAAV-AiE2419m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(378..1169,1170..1238))
                     /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     1001..1020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1239..1343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1411..1429
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1467..1489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1589..1608
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1625..2080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1762..1783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1974..1993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2155..2295
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2166..2295
                     /label=AAV2 ITR
     polyA_signal    2413..2524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2711..2731
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2824..3534,3535..3537,3538..3540))
                     /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(2866..2887)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3215..3234
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3474..3495
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3537..3546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4434..4563
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4625..5006,1..207))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4705..4724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
       61 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
      961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1381 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     1441 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1501 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1561 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1621 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1681 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     1741 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     1801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     1861 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     1921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     1981 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     2161 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2221 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     2281 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     2341 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     2401 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2461 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2521 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     2581 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     2641 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     2701 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     2761 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     2821 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     2881 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     2941 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     3001 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     3061 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     3121 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     3181 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     3241 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     3301 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     3361 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     3421 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     3481 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     3541 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     3601 agatggctct gccctgactt ttatgcccag cccgagctcc gtcctgtctc cacaacaaca
     3661 aaataagcac attaactgtc cctgcaggct gcccaagttg atataatcct gtctccattt
     3721 ctgactggtt ctataacctt gaacaaggta gttagcatct ctagcttggt ttcttccttg
     3781 gcgtaatgac atacaattgt attgccttca caggttgtga gaaggcaatg agctaatgca
     3841 tataaaactc tcccactgta tctggtacct aataagctca gagcagcaga tgttagctgt
     3901 taaaacagca ttggagaggc aacacagggc tgtctgctta taacaagtat aaggtcagta
     3961 tgcgatgagc acaagagcaa cactaaaaaa agaaatgatg tagtagccac aaaactgttt
     4021 cctccctctt cataacttca tgtatccttt ttggggcgtc actgttaggg tttcaattac
     4081 tgtgataaaa caccatgacc aaaagcaact tagcaaggca agaatttatc tcacttacac
     4141 tcttatataa ttcatcatgg atggaagtca ggaagaatct agaggcagga actaaagtaa
     4201 aggccacgaa gaagtgggtt atatgggctt gctcctcgtg acttgctcag cctgctttct
     4261 tttagcactc aagagcagca gctcaggagt ggcacgtgcc gagcaaactc tgtcctttcc
     4321 catcaactgt cagtgaagaa aatgtaccac aggcttgccc agaagccaat gtggtagggc
     4381 attttctcaa ttaagggtct ctcgattatt actcttacaa cgcgtgcggc cgcaggaacc
     4441 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4501 accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4561 aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4981 ctggtaacag gattagcaga gcgagg
//
