LOCUS       sequence-461840-        4959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0226h 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 4959)
  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 4959)
  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: AiP12311 - pAAV-AiE0226h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4959
                     /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      971..980
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(977..979,980..982,983..1693)
                     /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(1022..1043)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1283..1302)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1630..1651
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1786..1806)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1993..2104
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2222..2362
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2222..2351
                     /label=AAV2 ITR
     rep_origin      2437..2892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2524..2543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2734..2755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2909..2928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3028..3050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3088..3106)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3174..3278
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3279..3347,3348..4139)
                     /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(3497..3516)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4310..4898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4799..4818
                     /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 gcgtcacatt tctcatgaag ctgaggtagc ccagcattct
      181 ctactttaga aatctggatg tattttttcc tcatgggtct gtaattgagt ccgtaattga
      241 catttgtaca cttttaagat aaatctagcc tgtcttcaag tatattttag atctagtgaa
      301 atacagagta ctatagatta gatattctga atatcatctg tttgttattt agtaatgtga
      361 tatgccttgt gtacccactt ttgtatttac cattattgca aaatcaatca aatgtgagca
      421 aaagcaaagg caatttgaat gactcaaaat tgaaatttga gctgccaaat caagcagtaa
      481 aacaattttt accagctcta ttgattgtta gaaagataag ttataaactt tatttcaagt
      541 aaatttctaa aagatctggg aatgtgatta ttccaaggca gatggcgaag acctttattt
      601 ccactgatta ttcacagatg caaattatta tgcaactgga agcatactaa gatattgcaa
      661 agatgttctg acattagtca tctgctgcct ttgttacttt ggtgtcaatt ttcttattct
      721 ttccaaagga agatccttac agtttgtatt ctttcacagc tgggaaatga tcagttgaga
      781 attattcaaa cacaccaatc tgttaaccgt acttcttccc agataatgca atattttgca
      841 gggtgacagg caaaaagtgg tcatttttta cttcatagag ctcgggctgg gcataaaagt
      901 cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc
      961 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1021 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1081 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     1141 cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta
     1201 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1261 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1321 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1381 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     1441 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     1501 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1561 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
     1621 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1681 cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc
     1741 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1801 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1861 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     1921 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg
     1981 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2041 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2101 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2161 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     2221 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2281 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2341 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2401 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2461 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2521 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2581 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2641 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2701 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2761 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     2821 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2881 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2941 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3001 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3061 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3121 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3181 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3241 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3301 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3361 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3421 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3481 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3541 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3601 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3661 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3721 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3781 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3841 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3901 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3961 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4021 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4081 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4141 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4201 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4261 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4321 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4381 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4441 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4501 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4561 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4621 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4681 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4741 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4801 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4861 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4921 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
