LOCUS       sequence-470694-        5158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1016h 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 5158)
  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 5158)
  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: AiP13472 -
            pAAV-AiE1016h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(143..934,935..1003))
                     /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     766..785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1004..1108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1176..1194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1232..1254)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1354..1373
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1390..1845)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1527..1548)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1739..1758
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1920..2060
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1931..2060
                     /label=AAV2 ITR
     polyA_signal    2178..2289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2476..2496
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2583..2960)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(2991..3056)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3069..3125)
                     /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(3135..3845,3846..3848,3849..3851))
                     /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(3177..3198)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3526..3545
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3848..3857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4351..4480
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4542..5130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4622..4641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
       61 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
      121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
      181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
      241 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
      301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
      361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
      421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
      481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
      541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
      601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
      661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
      721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
      781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
      841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
      901 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
      961 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     1021 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1081 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     1141 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     1201 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1261 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1321 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     1381 tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1441 tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1501 agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1561 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1621 catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1681 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1741 ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg
     1801 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg
     1861 acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc
     1921 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     1981 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     2041 ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat
     2101 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     2161 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     2221 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     2281 aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct
     2341 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat
     2401 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     2461 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     2521 tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgcct
     2581 cattacttag cgctggtgta cttggtgatg gccttagtac cctcggacac ggcgtgcttg
     2641 gccaactccc caggcagcag caggcgcacg gccgtctgga tctcccggga ggtgatggtc
     2701 gagcgcttgt tgtaatgcgc caggcgggaa gcctctcctg cgatgcgctc gaaaatgtcg
     2761 ttcacaaacg aattcatgat gcccatggcc ttggacgaaa tgccggtgtc agggtggacc
     2821 tgcttcagaa ccttgtacac atagatggaa tagctctcct tgcggctgcg cttgcgcttc
     2881 ttgccgcctt tcttctgcgc cttagtcacc gccttcttgg agcccttttt cggggcggga
     2941 gcagacttcg ctggctctgg gcctccggag cctccagatc tgagtccgga cttgtcatcg
     3001 tcatccttgt aatcgatgtc atgatcttta taatctccgt catggtcttt gtagtctcca
     3061 ctagcaccag gccctggatt ctcctctaca tcccctgcct gtttcaacag tgagaagttg
     3121 gtagcaccag agcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     3181 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg
     3241 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     3301 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg
     3361 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg
     3421 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     3481 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     3541 tcgtctttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     3601 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg
     3661 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg
     3721 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     3781 ttgacatcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     3841 ttggagacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa
     3901 atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc agaaaacttg
     3961 tcagccccct acttggacta tcagtctgtt tgctttcaaa tgccatgctt tggaaatatc
     4021 tagacaagct ggaacaagct gaaaggaaat acagctgtaa gagttttgct gcatgtgaaa
     4081 tacagcagta agagttttgc tgcatatgaa atagctgcag aaattgtgct gaggagctaa
     4141 gagaagattc aaatgactat aatcctcatc tccaagggcc atagcttgag acagatttga
     4201 gagtatttat gatcagtaat tttccataag gatatagtga ctgtgttcta agagtgcatt
     4261 actgactcgc cagaggacag tggaagccct ttcctcagct gtgggcctaa gagaactata
     4321 gctgctgtct ccctgaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
     4381 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4441 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
     4501 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4561 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4621 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4681 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4741 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4801 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4861 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4921 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4981 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5041 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5101 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggt
//
