LOCUS       sequence-470714-        5206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1020h 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 5206)
  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 5206)
  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: AiP13612 -
            pAAV-AiE1020h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(263..282)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     610..631
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             683..739
                     /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             752..817
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             848..1225
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1312..1332)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1519..1630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1748..1888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1748..1877
                     /label=AAV2 ITR
     rep_origin      1963..2418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2050..2069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2260..2281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2435..2454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2554..2576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2614..2632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2700..2804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2805..2873,2874..3665)
                     /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     complement(3023..3042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3836..4424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4325..4344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4486..4615
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      5157..5166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5163..5165,5166..5168,5169..5206,1..673)
                     /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"
ORIGIN
        1 cctggtcgag ctggacggcg atgtcaacgg ccacaagttc agcgtgtccg gcgagggcga
       61 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
      121 cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta
      181 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      241 ggagcgcacc atcttcttca aagacgacgg caactacaag acccgcgccg aggtgaagtt
      301 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      361 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
      421 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
      481 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      541 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
      601 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      661 cgagctgtac aagggctctg gtgctaccaa cttctcactg ttgaaacagg caggggatgt
      721 agaggagaat ccagggcctg gtgctagtgg agactacaaa gaccatgacg gagattataa
      781 agatcatgac atcgattaca aggatgacga tgacaagtcc ggactcagat ctggaggctc
      841 cggaggccca gagccagcga agtctgctcc cgccccgaaa aagggctcca agaaggcggt
      901 gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg agagctattc
      961 catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt cgtccaaggc
     1021 catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag gagaggcttc
     1081 ccgcctggcg cattacaaca agcgctcgac catcacctcc cgggagatcc agacggccgt
     1141 gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta ctaaggccat
     1201 caccaagtac accagcgcta agtaatgagg cgcgccgcgg ccgcgaattc gatatcataa
     1261 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1321 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1381 ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat
     1441 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1501 ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1561 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1621 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1681 acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag
     1741 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     1801 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     1861 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     1921 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     1981 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     2041 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2101 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2161 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2221 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2281 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     2341 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2401 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2461 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2521 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2581 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     2641 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2701 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2761 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2821 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2881 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2941 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3001 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3061 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3121 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3181 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3241 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3301 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3361 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3421 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3481 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3541 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3601 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3661 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3721 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3781 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3841 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3901 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3961 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4021 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4081 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4141 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4201 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4261 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4321 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4381 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4441 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
     4501 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
     4561 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     4621 cgcacgcgta ggggaagctt tcatgccaag tggaaacaag acaaaataaa tgcaccgaat
     4681 ccgatttcaa aagctgtttg aaaactctgt ccggaattgc tggaaagggc cccagcatcc
     4741 acaggaaatg tatccctgcc aagccctgtg ctactgaact ctgtggcccc cactcctgcc
     4801 tcccggtgac ttgtttagaa ctacttcaga tcggaacaaa tcacagccaa ctgcagtcgc
     4861 tgtctctagc ccttcaatca ctcctaatca gaccgtgtat tttagcaaca gaatcttccc
     4921 tggctgagga gctcattcat ctgccttcca ccagctcgac accctttgaa tgggagatga
     4981 ggggaagcca cctgcttctc tggcagccaa gggaagctgg gaggaggcag gaattgccag
     5041 ccctccctct ctgatagatc accgagctcg ggctgggcat aaaagtcagg gcagagccat
     5101 ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca
     5161 ccatggtctc caagggcgag gagctgttca ccggggtggt gcccat
//
