LOCUS       sequence-461893-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0066h 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 4701)
  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 4701)
  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: AiP11410 - pAAV-AiE0066h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4701
                     /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      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(719..721,722..724,725..1435)
                     /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(764..785)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1025..1044)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1372..1393
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1528..1548)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1735..1846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1964..2104
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1964..2093
                     /label=AAV2 ITR
     rep_origin      2179..2634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2266..2285)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2476..2497
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2651..2670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2770..2792
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2830..2848)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2916..3020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3021..3089,3090..3881)
                     /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(3239..3258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4052..4640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4541..4560
                     /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 gcgtcatacc gatcgccctc actacactct gctttatata
      181 catattccct ctcggatcca tccatcactt ctttcaagat aataaaacaa ttcaatccca
      241 tcaaatcccc tgcaaccgct cccttgggag ggatgtttta tctctacttc tccgaaccca
      301 gggaggattt tacttgtttg aagggggcgg agaatggagc ggagtcctat ttatttccat
      361 tgatgactcc cctggttctc acaggcgctc agggaccggg tgggaggagg agaggaggag
      421 gtgcggagat tccctccgtc agaaaagaaa aagggttcag gaagcgaggc agcaggagga
      481 ggggtggagg ggggaaagag agggtagtgt gtttggggaa agcagcaact aaatgaaaaa
      541 ataaactctt ctttgtgatt tagcgggatc gacagatgag aagcaaattt atcaacccaa
      601 caaagttgct aaaaagaacg agctcgggct gggcataaaa gtcagggcag agccatctat
      661 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
      721 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      781 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      841 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      901 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      961 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1021 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1081 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1141 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1201 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1261 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1321 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1381 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt
     1441 cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg
     1501 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1561 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1621 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1681 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     1741 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1801 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1861 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1921 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     1981 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2041 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2101 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2161 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2221 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2281 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2341 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2401 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2461 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     2521 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     2581 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     2641 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     2701 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2761 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2821 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2881 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2941 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3001 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3061 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3121 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3181 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3241 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3301 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3361 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3421 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3481 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3541 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3601 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3661 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3721 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3781 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3841 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3901 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3961 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4021 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4081 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4141 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4201 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4261 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4321 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4381 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4441 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4501 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4561 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4621 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4681 gctggccttt tgctcacatg t
//
