LOCUS       sequence-459686-        4854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0475m 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 4854)
  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 4854)
  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: AiP12320 - pAAV-AiE0475m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4854
                     /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      866..875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(872..874,875..877,878..1588)
                     /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(917..938)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1178..1197)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1525..1546
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1681..1701)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1888..1999
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2117..2257
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2117..2246
                     /label=AAV2 ITR
     rep_origin      2332..2787
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2629..2650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2804..2823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2923..2945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2983..3001)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3069..3173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3174..3242,3243..4034)
                     /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(3392..3411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4205..4793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4694..4713
                     /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 gcgttatctt agagtgggaa gatttgagaa gtgccatggt
      181 taatatgact gactttttat tcttatttct tttaatttca tggttctaaa tccgaattta
      241 atcatagtac ccagaaaagc agaggtgtag aggttcacag tgggagttgt aatctagccc
      301 tattcatttt gacctcaaaa cccaaattat ttataacaaa ttatttccta ttctttcctt
      361 cactattcag gaacatctgt ccaccactta catgatcact tatcttgcta ttgtgtcatt
      421 ttgatgaaaa agaatttttt ctaaatatct aaatacaagg ccccatatta acagtgcttt
      481 ttaaatcccc acagatgtgg gagatgaccc ctttccatcc ctgaagattg taattgggcc
      541 agtctttagt acagtttgtt ccaataaaga gatacaattt tattcattaa tttgtgtatt
      601 catttagcaa atcactttag agtcttatta tatcaggatt ttggggtcta ttttagtata
      661 tctttttgta tttcttggaa cctctccaat tattctagac tctttcaaag gttggtgatc
      721 aatattagac attattatga aaagaatctt acttgctaaa agggttagat gtgagctcgg
      781 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
      841 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
      901 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      961 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1021 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     1081 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1141 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1201 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1261 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1321 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1381 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1441 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     1501 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1561 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     1621 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     1801 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1861 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     1921 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1981 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2041 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2101 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2161 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2221 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2281 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2341 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2401 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2461 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2521 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2581 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2641 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2701 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2761 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2821 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2881 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2941 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3001 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3061 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3121 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3181 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3241 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3301 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3361 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3421 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3481 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3541 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3601 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3661 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3721 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3781 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3841 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3901 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3961 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4021 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4081 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4141 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4201 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4261 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4321 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4381 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4441 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4501 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4561 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4621 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4681 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4741 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4801 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
