LOCUS       sequence-432549-        4887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2373m 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 4887)
  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 4887)
  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: AiP1698 - pAAV-AiE2373m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   166..306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   177..306
                     /label=AAV2 ITR
     polyA_signal    424..535
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     722..742
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(835..1545,1546..1548,1549..1551))
                     /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(877..898)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1226..1245
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1485..1506
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1548..1557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2326..2455
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2517..3105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2597..2616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3276..4067,4068..4136))
                     /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     3899..3918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4137..4241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4309..4327
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4365..4387)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4487..4506
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4523..4887,1..91))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4660..4681)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(4872..4887,1..4)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
       61 gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg
      121 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc
      181 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
      241 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      301 gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc
      361 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
      421 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
      481 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgaa
      541 gatctctcga gccacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc
      601 aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac caggatttat acaaggagga
      661 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
      721 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
      781 atccagaggt tgattatgat atcgaattcg cggccgcggc gcgccgtcga cttacttgta
      841 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
      901 atcgcgcttc tcgttggggt ctttgctcag cttggactgg tagctcaggt agtggttgtc
      961 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     1021 cacgccgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg
     1081 cttgtcggcg gtgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag
     1141 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     1201 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     1261 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     1321 gtcggggtag cgggcgaagc actgcacgcc gtagcccagg gtggtcacga gggtgggcca
     1381 gggcacgggc agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc
     1441 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     1501 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac
     1561 cggtagcgct agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa tagatggctc
     1621 tgccctgact tttatgccca gcccgagctc cgacaaacac acccctactt gccctcggcc
     1681 tcctgtcctt ccgcagagga caagccgggt tagactgact gggccggtca cctctgggat
     1741 aggaaaccgc agatcctcgc agaactcatg tgtccccctc gtctccattc tgagggtccc
     1801 aagggtcaag ccaaacggtg ggcattccaa gacggactag agaggattag gacagcggat
     1861 ggatctttag gggcgcaagg ggtgcagaac cacaaacctg cctcgggccg ccgcacccgg
     1921 gagctaggcg atctttctat agatagtaac agcgaagcgg ctgtttacag caacacagcg
     1981 ccgccagggc cgtctccagg cgacggagcg gccgggcgca cggtccgccc atccaggcca
     2041 cccgccctcc ctgctcccta ggaacctctg cctccctggc caggctgcgt ccgctcccgc
     2101 cgactgacat caccctcggg ttcggggggc tgatcccggc tggctcctgc ctctgcgtta
     2161 gcctccattc ttagagatcc aaacccagcg ggagcgggga gggcagggag gcggggattc
     2221 gtggaaatag gccacttgct atttttgcat tccctgtcac aggcagaggt gaaaacagaa
     2281 tttcatcatt gtatcgatta ttactcttac aacgcgtgcg gccgcaggaa cccctagtga
     2341 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2401 tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg
     2461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4261 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4321 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4381 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4441 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4501 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
     4561 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
     4621 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
     4681 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4741 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     4801 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4861 gcgagaaagg aagggaagaa agcgaaa
//
