LOCUS       sequence-470685-        5215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1011h 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 5215)
  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 5215)
  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: AiP13468 -
            pAAV-AiE1011h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(172..963,964..1032))
                     /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     795..814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1033..1137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1205..1223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1261..1283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1383..1402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1419..1874)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1556..1577)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1768..1787
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1949..2089
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1960..2089
                     /label=AAV2 ITR
     polyA_signal    2207..2318
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2505..2525
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2612..2989)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3020..3085)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3098..3154)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(3164..3874,3875..3877,3878..3880))
                     /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(3206..3227)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3555..3574
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3877..3886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4437..4566
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4628..5215,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4708..4727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
       61 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     1201 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1261 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1321 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1381 tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt
     1441 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat
     1501 cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt
     1561 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1621 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     1681 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1741 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc
     1801 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1861 gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc
     1921 gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact
     1981 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2041 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcg
     2101 gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     2161 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     2221 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     2281 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct cgagccacgg
     2341 aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga
     2401 gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg aaagccatac
     2461 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     2521 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat
     2581 gatatcgaat tcgcggccgc ggcgcgcctc attacttagc gctggtgtac ttggtgatgg
     2641 ccttagtacc ctcggacacg gcgtgcttgg ccaactcccc aggcagcagc aggcgcacgg
     2701 ccgtctggat ctcccgggag gtgatggtcg agcgcttgtt gtaatgcgcc aggcgggaag
     2761 cctctcctgc gatgcgctcg aaaatgtcgt tcacaaacga attcatgatg cccatggcct
     2821 tggacgaaat gccggtgtca gggtggacct gcttcagaac cttgtacaca tagatggaat
     2881 agctctcctt gcggctgcgc ttgcgcttct tgccgccttt cttctgcgcc ttagtcaccg
     2941 ccttcttgga gccctttttc ggggcgggag cagacttcgc tggctctggg cctccggagc
     3001 ctccagatct gagtccggac ttgtcatcgt catccttgta atcgatgtca tgatctttat
     3061 aatctccgtc atggtctttg tagtctccac tagcaccagg ccctggattc tcctctacat
     3121 cccctgcctg tttcaacagt gagaagttgg tagcaccaga gcccttgtac agctcgtcca
     3181 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     3241 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     3301 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     3361 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3421 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3481 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3541 tcacctcggc gcgggtcttg tagttgccgt cgtctttgaa gaagatggtg cgctcctgga
     3601 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3661 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     3721 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3781 cctcgccgga cacgctgaac ttgtggccgt tgacatcgcc gtccagctcg accaggatgg
     3841 gcaccacccc ggtgaacagc tcctcgccct tggagaccat ggtggcgacc ggtagcgcta
     3901 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     3961 ttatgcccag cccgagctct gtaactatgt agtatgaagt tgccttaaac tagaaatgtg
     4021 ccttctattc tgtggattgg caccaccagt tccatgaatt tagcattttc ctttaggaaa
     4081 tcaaaagcta tgcagattac tcattgaaat ttagagagtg aaaggcattt ttcatcagca
     4141 catatcatat aattggcaca aaggcttgga aatgaagaat tgcctgttag ttctgagaca
     4201 gtgtttatta tgcaaaatat aaataccaga attggggttg gggcacatcc aacaccctgt
     4261 gttcaacaag attaaaattt ggggggacca ggattatgat ttttttcatg tttgtttttg
     4321 gcccactttc tccagtacca aaatatggaa ggccaaacat ttgtatatac ttgttaactg
     4381 atgcattgca ttgtatgtcg taacaagtac agatggcagt gtacgcgtgc ggccgcagga
     4441 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4501 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4561 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5041 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctca
//
