LOCUS       sequence-461845-        4642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0022h 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 4642)
  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 4642)
  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: AiP12376 - pAAV-AiE0022h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(660..662,663..665,666..1376)
                     /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(705..726)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(966..985)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1313..1334
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1469..1489)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1676..1787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1905..2045
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1905..2034
                     /label=AAV2 ITR
     rep_origin      2120..2575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2207..2226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2417..2438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2592..2611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2711..2733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2771..2789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2857..2961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2962..3030,3031..3822)
                     /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(3180..3199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3993..4581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4482..4501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagtcct gtggatacca tctgagcctg tgcccttttc
      181 tctctactct tgagcaagtg cccatcagga gccacatttg agtcaggacc tgccaagagc
      241 attttcatct catcctcagt tccccttttg tctggaacca ctcgctatga gtttctatga
      301 gatcgtagcc aggtttgacc aaaccactcc tgcacacatg ttgttttgaa agattagcac
      361 agaatccaac acaacatttc ccttggattc atctttcaag ccaacagaga aagagctatt
      421 tccaaaaaaa taacacatta ttaaccaatg aataagagcc aagaaaacaa tgtagccagc
      481 atatttattc caaacagctt gtttagaatg caaattgcaa atacatctat ttccacccgg
      541 ctctttgggg aaccatgctg gagctcgggc tgggcataaa agtcagggca gagccatcta
      601 ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca
      661 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      721 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      781 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      841 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
      901 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      961 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1021 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1081 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     1141 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     1201 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1261 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1321 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     1381 tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt
     1441 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1501 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1561 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1621 ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg
     1681 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1741 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1801 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1861 gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     1921 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     1981 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     2041 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2101 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2161 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     2221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     2341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     2461 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     2521 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     2581 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2641 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2701 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2761 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2821 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2881 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2941 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3001 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3061 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3121 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3181 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3241 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3301 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3361 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3421 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3481 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3541 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3601 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3661 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3721 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3781 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3841 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3901 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3961 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4021 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4081 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4141 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4201 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4261 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4321 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4381 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4441 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4501 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4561 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4621 tgctggcctt ttgctcacat gt
//
