LOCUS       sequence-470498-        4663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0565h 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 4663)
  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 4663)
  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: AiP12681 - pAAV-AiE0565h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   66..195
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(257..845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(337..356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1016..1807,1808..1876))
                     /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     1639..1658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1877..1981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2049..2067
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2105..2127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2227..2246
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2263..2718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2612..2631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2793..2933
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2804..2933
                     /label=AAV2 ITR
     polyA_signal    3051..3162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3349..3369
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3462..4172,4173..4175,4176..4178))
                     /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(3504..3525)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3853..3872
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4112..4133
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4175..4184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agatttaagt tgagttgtct tatcattgtc tttggtcttt aattgaggaa gacgcgtgcg
       61 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
      121 tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc
      181 gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1981 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     2041 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     2101 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2161 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     2221 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     2281 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     2341 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     2401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     2521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2641 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2701 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
     2761 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
     2821 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     2881 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     2941 ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     3001 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     3061 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     3121 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaagat ctctcgagcc
     3181 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     3241 atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa aatgaaagcc
     3301 atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa
     3361 ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga
     3421 ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg
     3481 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     3541 ttggggtctt tgctcagctt ggactggtag ctcaggtagt ggttgtcggg cagcagcacg
     3601 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc
     3661 tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg
     3721 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     3781 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     3841 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     3901 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     3961 gcgaagcact gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc
     4021 ttgccggtgg tgcagatcag cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     4081 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     4141 accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc
     4201 ttcgaaagat ctggatccca gaagcaaatg taagcaatag atggctctgc cctgactttt
     4261 atgcccagcc cgagctcacc tcctatgtgc ttagttcagg ttttaagaag tgaatttctg
     4321 gaaacaacgc caggttgtgt aagatggatg aaacgtttgc atgctgatgt ctcttggtaa
     4381 agtccctgcg cttagcctac cctttggtat taatgcggga agtatttttg tctcgtagag
     4441 ctctggctgg agtggctgca tgacgagatc agcatggccc aggatggcct ggacagagag
     4501 cacgtgtatg acctctttga gaaagccgtg aaggattaca tttgtaagtt cccgtggact
     4561 ttttttggat atctggcaat gtcatatata tcagtctgct gttgaggact gtattatgag
     4621 atgttgctac ccagaaaagg gaacgttagt ttcacaatta gca
//
