LOCUS       sequence-470697-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1008h 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 5455)
  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 5455)
  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: AiP13490 -
            pAAV-AiE1008h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   462..591
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(653..1241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(733..752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1412..2203,2204..2272))
                     /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     2035..2054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2273..2377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2445..2463
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2501..2523)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2623..2642
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2659..3114)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2796..2817)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3008..3027
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3189..3329
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3200..3329
                     /label=AAV2 ITR
     polyA_signal    3447..3558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3745..3765
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3852..4229)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(4260..4325)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4338..4394)
                     /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(4404..5114,5115..5117,5118..5120))
                     /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(4446..4467)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4795..4814
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      5117..5126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgcagctgga atcagggtgt ctgacaagag gcagctgtca cttcacattt tgcaaaatat
       61 ttccacaaaa attgccccaa aatgcaaaaa tctttcatta ggaagagctg attcatggaa
      121 tttataatag gaatcctgta tggctgcata attcagtaga ggaaaataga aaagaataaa
      181 caagtattgg tctctccagg ccattccttt tgaagccaag tcagggcaca gagcactatg
      241 gacactgcat gaccagcctg gttgaaggca atgttggaga taaactttcc cacacagaaa
      301 cccttctcag ccctgagtct ctgtaaaaat gcacagctct tgcatgtaca tagagacttc
      361 tcttccccag gcagaggagt caacgtgaaa gccaaagcca aagcacacta gcccttcaga
      421 gatcaaaaac cctgtgacac tctcagaacg cgtgcggccg caggaacccc tagtgatgga
      481 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
      541 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag
      601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1081 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1321 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1381 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1441 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1501 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1561 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1621 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1681 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1741 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1801 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1861 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1921 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1981 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2041 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2101 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2161 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2221 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2341 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2401 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2461 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2521 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2581 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     2641 tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     2701 tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat
     2761 caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat
     2821 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2881 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2941 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3001 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
     3061 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
     3121 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     3181 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     3241 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     3301 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
     3361 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     3421 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     3481 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     3541 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
     3601 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
     3661 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     3721 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     3781 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
     3841 ggcgcgcctc attacttagc gctggtgtac ttggtgatgg ccttagtacc ctcggacacg
     3901 gcgtgcttgg ccaactcccc aggcagcagc aggcgcacgg ccgtctggat ctcccgggag
     3961 gtgatggtcg agcgcttgtt gtaatgcgcc aggcgggaag cctctcctgc gatgcgctcg
     4021 aaaatgtcgt tcacaaacga attcatgatg cccatggcct tggacgaaat gccggtgtca
     4081 gggtggacct gcttcagaac cttgtacaca tagatggaat agctctcctt gcggctgcgc
     4141 ttgcgcttct tgccgccttt cttctgcgcc ttagtcaccg ccttcttgga gccctttttc
     4201 ggggcgggag cagacttcgc tggctctggg cctccggagc ctccagatct gagtccggac
     4261 ttgtcatcgt catccttgta atcgatgtca tgatctttat aatctccgtc atggtctttg
     4321 tagtctccac tagcaccagg ccctggattc tcctctacat cccctgcctg tttcaacagt
     4381 gagaagttgg tagcaccaga gcccttgtac agctcgtcca tgccgagagt gatcccggcg
     4441 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc
     4501 ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     4561 gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc
     4621 ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg
     4681 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     4741 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     4801 tagttgccgt cgtctttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     4861 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg
     4921 tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc
     4981 agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     5041 ttgtggccgt tgacatcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     5101 tcctcgccct tggagaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc
     5161 cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctct
     5221 ggacaaagtg ctcagttgac attcattaaa ctgatcatcg gtaccaagaa attacaaata
     5281 gatgcgtaaa aatctggaaa cagtctttct cagcaaccat tccagaatta caaactttct
     5341 ctagcatgac cgtaggatct aaatgcagtt ccagtggaat taaatgtatg ggttgttttg
     5401 tgtctatagc atgactcctt cctgtctgag aactttctgg aacccaaatt gctag
//
