LOCUS       sequence-470700-        5432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1013h 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 5432)
  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 5432)
  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: AiP13491 -
            pAAV-AiE1013h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    151..262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     449..469
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(556..933)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(964..1029)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1042..1098)
                     /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(1108..1818,1819..1821,1822..1824))
                     /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(1150..1171)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1499..1518
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      1821..1830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2598..2727
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2789..3377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2869..2888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3548..4339,4340..4408))
                     /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     4171..4190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4409..4513)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4581..4599
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4637..4659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4759..4778
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4795..5250)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4932..4953)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5144..5163
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5325..5432,1..33)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5336..5432,1..33)
                     /label=AAV2 ITR
ORIGIN
        1 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
       61 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
      121 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
      181 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
      241 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
      301 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
      361 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
      421 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
      481 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
      541 ccgcggcgcg cctcattact tagcgctggt gtacttggtg atggccttag taccctcgga
      601 cacggcgtgc ttggccaact ccccaggcag cagcaggcgc acggccgtct ggatctcccg
      661 ggaggtgatg gtcgagcgct tgttgtaatg cgccaggcgg gaagcctctc ctgcgatgcg
      721 ctcgaaaatg tcgttcacaa acgaattcat gatgcccatg gccttggacg aaatgccggt
      781 gtcagggtgg acctgcttca gaaccttgta cacatagatg gaatagctct ccttgcggct
      841 gcgcttgcgc ttcttgccgc ctttcttctg cgccttagtc accgccttct tggagccctt
      901 tttcggggcg ggagcagact tcgctggctc tgggcctccg gagcctccag atctgagtcc
      961 ggacttgtca tcgtcatcct tgtaatcgat gtcatgatct ttataatctc cgtcatggtc
     1021 tttgtagtct ccactagcac caggccctgg attctcctct acatcccctg cctgtttcaa
     1081 cagtgagaag ttggtagcac cagagccctt gtacagctcg tccatgccga gagtgatccc
     1141 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     1201 cagcttggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     1261 gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg
     1321 gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg
     1381 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1441 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1501 cttgtagttg ccgtcgtctt tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     1561 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac
     1621 gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     1681 gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     1741 gaacttgtgg ccgttgacat cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1801 cagctcctcg cccttggaga ccatggtggc gaccggtagc gctagcttcg aaagatctgg
     1861 atcccagaag caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag
     1921 ctcacctgaa tctctcctgc ctcaaaaatt tcattccctg atgatgatgt gaaaataggg
     1981 agggagatag tttgacaggt cagatcaaca ggctctctcg atttctggca gaaatattcc
     2041 cctagtgaca tttgctccca tcctgtggcc cctacaggct gcttcctagg gctgcccagc
     2101 aaatcaaggc agggctctct ccaaggaatt gtgagacctg cccccatagg tcaggtgtcc
     2161 ctgcagccct tggcctcctg ctgctgtcgc caccctgcca ggccctgtga tgggagcaga
     2221 gcgcactgcg tcagccattg ggtggccaca cagcctatgc agccaggaag gtggcgagct
     2281 tgagatgtga gaactggctg ctcagactca gggatgctcc ctgcagacta gcaaatgagc
     2341 ccggaaggat gcgcagactc tgcgaacaac cagcctcagg caactctgac cctgtgctca
     2401 tgatcagttg ggaagaggcc aaaagtcggt ccaaactgct ctttatgtcc tggcaacact
     2461 cacactaggg ctgcctcatc tactgacgct taattccctt tgtctgatgg tttggtaaaa
     2521 gattcaggac cacatttaag tcctactctt ccccagccag caccaccacc actagcacca
     2581 aatacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2641 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     2701 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
     2761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3841 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4141 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4501 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4561 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4621 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4681 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4741 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     4801 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4861 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     4921 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4981 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     5041 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5101 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5161 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5221 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     5281 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     5341 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     5401 cccggcctca gtgagcgagc gagcgcgcag ag
//
