LOCUS       sequence-460620-        4554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0536h 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 4554)
  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 4554)
  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: AiP12471 - pAAV-AiE0536h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4554
                     /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      566..575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(572..574,575..577,578..1288)
                     /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(617..638)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(878..897)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1225..1246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1381..1401)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1588..1699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1817..1957
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1817..1946
                     /label=AAV2 ITR
     rep_origin      2032..2487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2119..2138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2329..2350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2504..2523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2623..2645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2683..2701)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2769..2873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2874..2942,2943..3734)
                     /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(3092..3111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3905..4493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4394..4413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtccagga gcaaagcagc gcgccaggct ggagctgttc
      181 cgtcattacc gccacgtctg cgcttcggtt gtgagaaccg attcgttacc ctaacctctt
      241 aggactaaga gccagtcttc tggtggcgct cgctttcact tactttttat ttccctttct
      301 attcccgcga gctaggggga gaaagtgtaa atcccgcccg cttctccagg gctgaaaatg
      361 gagaggctga ggcggcttgg agagtgcgca ctgcgcatgc tccgcagagc atccccgcag
      421 gtcaaccggg gcaagggaga gacatgaaag cctgtggctt ctcctttgaa gagagctcgg
      481 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
      541 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
      601 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      661 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
      721 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
      781 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      841 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      901 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      961 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1021 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1081 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1141 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     1201 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1261 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     1321 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1381 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1441 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     1501 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1561 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     1621 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1681 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1741 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     1801 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     1861 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     1921 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     1981 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2041 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2101 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2161 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2221 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2281 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2341 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2401 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2461 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2521 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2581 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2641 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2701 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2881 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2941 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3001 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3061 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3121 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3181 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3241 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3301 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3361 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3421 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3481 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3541 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3601 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3661 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3721 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3781 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3841 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3901 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3961 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4021 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4081 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4141 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4201 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4261 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4321 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4381 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4441 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4501 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
