LOCUS       sequence-470774-        5628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0363h 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 5628)
  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 5628)
  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: AiP14523 -
            pAAV-AiE0363h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    149..260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     447..467
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(554..931)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(962..1027)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1040..1096)
                     /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(1106..1816,1817..1819,1820..1822))
                     /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(1148..1169)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1497..1516
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      1819..1828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2679..2808
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    2815..2842
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2843..2870
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     rep_origin      complement(2927..3515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3007..3026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3686..4477,4478..4546))
                     /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     4309..4328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4547..4651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4719..4737
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4775..4797)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4897..4916
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4933..5388)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5070..5091)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5282..5301
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_feature    5457..5484
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5485..5512
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     repeat_region   join(5519..5628,1..31)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5530..5628,1..31)
                     /label=AAV2 ITR
ORIGIN
        1 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc
       61 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
      121 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
      181 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      241 ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca
      301 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa
      361 ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
      421 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
      481 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc
      541 gcggcgcgcc tcattactta gcgctggtgt acttggtgat ggccttagta ccctcggaca
      601 cggcgtgctt ggccaactcc ccaggcagca gcaggcgcac ggccgtctgg atctccctgg
      661 aggtgatggt cgagcgcttg ttgtaatgcg ccaggcggga agcctcacct gcgatgcgct
      721 cgaaaatgtc gttcacaaac gagttcatga tgcccatggc cttggacgaa atgccggtgt
      781 cagggtggac ctgcttcaga accttataca catagatgga atagctctcc ttgcggctgc
      841 gcttgcgctt cttgccgcct ttcttctgcg ccttagtcac cgccttcttg gagccctttt
      901 tcggggcggg agcagacttc gctggctctg ggcctccgga gcctccagat ctgagtccgg
      961 acttgtcatc gtcatccttg taatcgatgt catgatcttt ataatctccg tcatggtctt
     1021 tgtagtctcc actagcacca ggccctggat tctcctctac atcccctgcc tgtttcaaca
     1081 gtgagaagtt ggtagcacca gagcccttgt acagctcgtc catgccgaga gtgatcccgg
     1141 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     1201 gcttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     1261 gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga
     1321 tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc
     1381 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     1441 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     1501 tgtagttgcc gtcgtctttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     1561 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc
     1621 cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1681 tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     1741 acttgtggcc gttgacatcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     1801 gctcctcgcc cttggagacc atggtggcga ccggtagcgc tagcttcgaa agatctggat
     1861 cccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct
     1921 cttacggtat tcccttgtga agtgggcaag gcaagttaat agtatcccat tttgagaatg
     1981 ggaaaactga gccttggaga atagatttgt tgtgagaaaa tggagaccag agggtttgag
     2041 tgacttatcc aagactacac agctcctaag tggctaagtg tgggctcgaa tttagatcag
     2101 gtggctccaa gctcagggtt gcatcatccc actcagctgt gtctctgaag ggcactgcat
     2161 cagattcttc tccaccctaa tttgtgcgat gtttttctct ccatgagagc actgtactgt
     2221 gtttctacaa aatatgagtg tctttctgaa attggccaaa atctccgtca tccaaatgtt
     2281 gataaacaac acatgcaatg gcccacttgt tgctgcaccc cctcttctac tgtcctccca
     2341 aaggaagctt tgtcttaggg ctgctgacat cagtcctcag tgcccactga tgagataagg
     2401 ctctgggcag ctgcattcag tgcagcgacc atccagtgtc cttgcagcac gtgttggtct
     2461 ggtgctagcg tgaagagcag tggttaggtc ccttgagcaa gaggaccctg acctaagagg
     2521 cccagacgtc cctgtgttcc accgtcatcc cagaagcaga ggtgcacagt gctgtcagcc
     2581 tgggtgctct gattcatcca cagagactac tggagttgct tggaagcctt gcagagtggt
     2641 ggaaagaggt tgtcaccggg acacacgcgt gcggccgcag gaacccctag tgatggagtt
     2701 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2761 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgttatcag
     2821 cacacaatag tccattatac gcgcgtataa tgggcaattg tgtgctgata tgagcaaaag
     2881 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2941 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3001 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3061 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3121 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3361 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3661 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3721 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3781 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3841 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3901 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3961 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4021 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4081 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4141 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4201 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4261 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4321 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4381 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4441 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4501 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4561 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4621 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4681 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4741 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4801 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4861 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4921 gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     4981 aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag
     5041 aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5161 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5281 aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc
     5341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     5401 ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcatatc
     5461 agcacacaat agtccattat acgcgcgtat aatgggcaat tgtgtgctga taggcgcccc
     5521 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     5581 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagag
//
