LOCUS       sequence-461879-        5393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1017h 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 5393)
  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 5393)
  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: AiP13473 -
            pAAV-AiE1017h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5393
                     /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      859..868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(865..867,868..870,871..1581)
                     /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(1171..1190)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1518..1539
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1591..1647
                     /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             1660..1725
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1756..2133
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2220..2240)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2427..2538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2656..2796
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2656..2785
                     /label=AAV2 ITR
     rep_origin      2871..3326
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2958..2977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3168..3189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3343..3362)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3462..3484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3522..3540)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3608..3712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3713..3781,3782..4573)
                     /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(3931..3950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4744..5332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5233..5252
                     /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 gcgttgtgtg accggttcct aacaggtcac cattagtact
      181 ggtccgtggc ccgggtaatg gggacccctg ttgtaaacca tggatgtgag agagaacctg
      241 gcttgttcaa ggaactgaaa agaagagagg acttggtcag aatgtccttg tcatgtgggc
      301 cttatgagcc atgttaagcc gtttggactt ctgactaaaa gacatgggac atcactgaag
      361 ggttttgagc aggggaacaa cagcatattt ttgtgttaga aagattattt taaatgcaga
      421 gtgggcggga tgagagactg gaagttggtg gatgttgtag tcatatgaat gggagatgat
      481 gaatagcggg ccgaccagat gcagagcaag ggacagggag tgctggaatg aacaggaagt
      541 gttggctgat tggtgtgcag gttgagacag gaggcaagga tgaatgccag gcttctggct
      601 tgccagctaa gtccaggcgt tgctattctc tgagataagg gaccctgtct gaggtgaacc
      661 attcagattt ggagatactg agtttgacga ggttatgaga caccgaagtg agagtgtctg
      721 gtaggcaggt ggaagtatgg gcctggagct caggaggtac actaggagct cgggctgggc
      781 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      841 aagctagcgc taccggtcgc caccatggtc tccaagggcg aggagctgtt caccggggtg
      901 gtgcccatcc tggtcgagct ggacggcgat gtcaacggcc acaagttcag cgtgtccggc
      961 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     1021 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     1081 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1141 tacgtccagg agcgcaccat cttcttcaaa gacgacggca actacaagac ccgcgccgag
     1201 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1261 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1321 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1381 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1441 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1501 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1561 ggcatggacg agctgtacaa gggctctggt gctaccaact tctcactgtt gaaacaggca
     1621 ggggatgtag aggagaatcc agggcctggt gctagtggag actacaaaga ccatgacgga
     1681 gattataaag atcatgacat cgattacaag gatgacgatg acaagtccgg actcagatct
     1741 ggaggctccg gaggcccaga gccagcgaag tctgctcccg ccccgaaaaa gggctccaag
     1801 aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag
     1861 agctattcca tctatgtgta caaggttctg aagcaggtcc accctgacac cggcatttcg
     1921 tccaaggcca tgggcatcat gaattcgttt gtgaacgaca ttttcgagcg catcgcagga
     1981 gaggcttccc gcctggcgca ttacaacaag cgctcgacca tcacctcccg ggagatccag
     2041 acggccgtgc gcctgctgct gcctggggag ttggccaagc acgccgtgtc cgagggtact
     2101 aaggccatca ccaagtacac cagcgctaag taatgaggcg cgccgcggcc gcgaattcga
     2161 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2221 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2281 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     2341 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2401 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     2461 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2521 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2581 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2641 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2701 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2761 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2821 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2881 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2941 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3001 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3061 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3121 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3181 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3241 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3301 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3361 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3481 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3541 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
