LOCUS       sequence-460625-        5363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1391h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5363)
  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 5363)
  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: AiP14981 -
            pAAV-AiE1391h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      716..725
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(722..724,725..727,728..1438)
                     /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(1028..1047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1375..1396
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1448..1504
                     /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             1517..1582
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1613..1990
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2077..2097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2284..2395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2513..2653
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2513..2642
                     /label=AAV2 ITR
     misc_feature    2660..2687
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2688..2715
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2784..3239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2871..2890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3081..3102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3256..3275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3375..3397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3435..3453)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3521..3625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3626..3694,3695..4486)
                     /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(3844..3863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4657..5245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5146..5165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5302..5329
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5330..5357
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcactgt gatgcttcta agagctttaa acttgtaaat
      181 aacccctccc agaagcgcaa atctccatat cttaatatca tgaaagctgg aactgtaaaa
      241 taaaatctgt tagcagctga aatcttaatg aggttctaca aagaagaggc ttttctgcaa
      301 tgactcaatg catggaaaca gttgagtaat tataaatgca tgggctgaat gtgagttggc
      361 aaatataata ccacagaatg tacccagaac tagggttgtc ccagttcaaa tgagactata
      421 ttcctttacc agtacaggca aacaactaac ctgaaatgtc tttggctaag agataggtgt
      481 gagcccataa aattatgtcc cttgactcct catttacatg gtgacaatct gttctggccc
      541 accagcagcc tgttcaacaa caaagcactt cagaaagagc tctgcataat attggtttca
      601 tcagacataa tagaagccaa tggagctcgg gctgggcata aaagtcaggg cagagccatc
      661 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
      721 catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      781 cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      841 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
      901 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
      961 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1021 cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1081 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1141 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1201 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1261 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1321 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1381 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1441 ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg
     1501 gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc atgacatcga
     1561 ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc
     1621 agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa
     1681 gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtataa
     1741 ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa
     1801 ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta
     1861 caacaagcgc tcgaccatca cctccaggga gatccagacg gccgtgcgcc tgctgctgcc
     1921 tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag
     1981 cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     2041 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2101 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2161 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     2221 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     2281 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2341 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2401 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2461 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2521 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2581 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2641 agctgcctgc aggggcgcct atcagcacac aattgcccat tatacgcgcg tataatggac
     2701 tattgtgtgc tgatatgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2761 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3121 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3181 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3241 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3421 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3481 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3841 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4141 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4321 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4801 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5281 cttttgctgg ccttttgctc atatcagcac acaattgccc attatacgcg cgtataatgg
     5341 actattgtgt gctgataaca tgt
//
