LOCUS       sequence-470673-        5382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0979h 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 5382)
  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 5382)
  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: AiP13463 -
            pAAV-AiE0979h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(152..220,221..1012)
                     /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(370..389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1183..1771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1672..1691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1833..1962
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2680..2689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2686..2688,2689..2691,2692..3402)
                     /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(2992..3011)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3339..3360
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3412..3468
                     /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             3481..3546
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3577..3954
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4041..4061)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4248..4359
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4477..4617
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4477..4606
                     /label=AAV2 ITR
     rep_origin      4692..5147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4779..4798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4989..5010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5164..5183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5283..5305
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5343..5361)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
       61 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      121 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      181 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      241 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
      301 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
      361 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
      421 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
      481 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
      541 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
      601 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
      661 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
      721 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
      781 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
      841 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
      901 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
      961 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1021 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1081 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1141 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1201 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1261 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1321 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1381 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1441 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1501 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1561 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1621 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1681 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1741 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1801 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
     1861 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     1921 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtttcc
     1981 agcctgattc cctgacaagg tgtggcttct tattcttcaa gactccgttc tagggccacg
     2041 atgtggaggg agctgggtgg ctggctgcct gggcttcctc ccggctgctc atttctgcct
     2101 gtgggacagc catccccatg tgtcgctgtg gtcttctctc tggtcctgct ggcctcctgg
     2161 accaagctgc aagcatctgg aaggcagaga ctgaacgtga gccacatttc tgtcctcaac
     2221 atctcctagg atgcctggtg cagagctggc actcaattca gacctgtaga caagcatgac
     2281 accatctatg tttaccaaag actgggataa aaccgcaact cagcatttga ggtccaccct
     2341 gagcactgga ggttctccaa gtacttcctc cttctgcctc tcagtacaga tcgcgtcttc
     2401 taggtgtgct gtgcacattc catcacccac cttctggctc atcgcacttg ctctgcttcc
     2461 agtgagtctc accttcccaa atcctactca tgcatccagg cctatctcaa gccttcctca
     2521 tccactaatg cagacttctc tgattccgtg agcctttcct gccctcttct tgcttggcca
     2581 cccatagagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
     2641 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt ctccaagggc
     2701 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga tgtcaacggc
     2761 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2821 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2881 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2941 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa agacgacggc
     3001 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3061 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3121 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     3181 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     3241 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     3301 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3361 accgccgccg ggatcactct cggcatggac gagctgtaca agggctctgg tgctaccaac
     3421 ttctcactgt tgaaacaggc aggggatgta gaggagaatc cagggcctgg tgctagtgga
     3481 gactacaaag accatgacgg agattataaa gatcatgaca tcgattacaa ggatgacgat
     3541 gacaagtccg gactcagatc tggaggctcc ggaggcccag agccagcgaa gtctgctccc
     3601 gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag
     3661 cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc
     3721 caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac
     3781 attttcgagc gcatcgcagg agaggcttcc cgcctggcgc attacaacaa gcgctcgacc
     3841 atcacctccc gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag
     3901 cacgccgtgt ccgagggtac taaggccatc accaagtaca ccagcgctaa gtaatgaggc
     3961 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     4021 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4081 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4141 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4201 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     4261 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4321 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4381 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4441 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4501 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4561 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4621 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4681 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4741 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     5041 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     5101 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5161 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5221 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5281 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5341 gggcctcgtg atacgcctat ttttataggt taatgtcatg at
//
