LOCUS       sequence-470679-        5290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1009h 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 5290)
  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 5290)
  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: AiP13466 -
            pAAV-AiE1009h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   379..508
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(570..1158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(650..669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1329..2120,2121..2189))
                     /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     1952..1971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2190..2294)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2362..2380
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2418..2440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2540..2559
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2576..3031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2713..2734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2925..2944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3106..3246
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3117..3246
                     /label=AAV2 ITR
     polyA_signal    3364..3475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3662..3682
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3769..4146)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(4177..4242)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4255..4311)
                     /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(4321..5031,5032..5034,5035..5037))
                     /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(4363..4384)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4712..4731
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      5034..5043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggaatgggaa cattgaattg caagatgttg ttgattatga ccaggagtct gattcatatg
       61 ctaatcagtg aaaacagtaa caaatactcc aaattgatgt accatacaat ttatcatggc
      121 tttgggatgt attttggcac acagtgtact acagtgattc taagatgaac attttaattt
      181 taaagtcttt gcagtcattg atggcatgtt gtaatttaat ggcacataaa acaatggtgc
      241 atttttaatc aatgacagat tcgatgaaat atggttcatg tagattttaa tgtaactgac
      301 agcctggaaa ttcaaatcat tttgaaccaa ataaagtatg catgaacatc gataattatt
      361 gccaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
      421 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg
      481 agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg
      541 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      601 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      661 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      721 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      781 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      841 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      901 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      961 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1021 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1081 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1141 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1201 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1261 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1321 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1381 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1441 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1501 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1561 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1621 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1681 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1741 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1801 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1861 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1921 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     1981 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2041 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2101 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2161 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2221 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2281 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2341 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     2401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2521 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaaatt
     2581 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2641 aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagatagag
     2701 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2761 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     2821 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2881 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2941 ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3001 gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg
     3061 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc
     3121 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
     3181 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     3241 gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc
     3301 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     3361 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     3421 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgaa
     3481 gatctctcga gccacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc
     3541 aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac caggatttat acaaggagga
     3601 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     3661 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     3721 atccagaggt tgattatgat atcgaattcg cggccgcggc gcgcctcatt acttagcgct
     3781 ggtgtacttg gtgatggcct tagtaccctc ggacacggcg tgcttggcca actccccagg
     3841 cagcagcagg cgcacggccg tctggatctc ccgggaggtg atggtcgagc gcttgttgta
     3901 atgcgccagg cgggaagcct ctcctgcgat gcgctcgaaa atgtcgttca caaacgaatt
     3961 catgatgccc atggccttgg acgaaatgcc ggtgtcaggg tggacctgct tcagaacctt
     4021 gtacacatag atggaatagc tctccttgcg gctgcgcttg cgcttcttgc cgcctttctt
     4081 ctgcgcctta gtcaccgcct tcttggagcc ctttttcggg gcgggagcag acttcgctgg
     4141 ctctgggcct ccggagcctc cagatctgag tccggacttg tcatcgtcat ccttgtaatc
     4201 gatgtcatga tctttataat ctccgtcatg gtctttgtag tctccactag caccaggccc
     4261 tggattctcc tctacatccc ctgcctgttt caacagtgag aagttggtag caccagagcc
     4321 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     4381 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc tcaggtagtg
     4441 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     4501 gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt
     4561 cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     4621 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     4681 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ctttgaagaa
     4741 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     4801 catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg tcacgagggt
     4861 gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca gcttgccgta
     4921 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttga catcgccgtc
     4981 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgg agaccatggt
     5041 ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt aagcaataga
     5101 tggctctgcc ctgactttta tgcccagccc gagctcagca gtggagaaag agctctgttt
     5161 ctgtcttcaa gagggcatac tgcatatttc ccagagacaa aagggagcat cgaacccaat
     5221 gccagtttgc taacagagcc ttccatatgc aagcaccaga gttctatgct tcaccaggtt
     5281 ggtcatttct
//
