LOCUS       sequence-449775-        4892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0647m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L5_ET neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  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 4892)
  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: AiP12847 - pAAV-AiE0647m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      904..913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(910..912,913..915,916..1626)
                     /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(955..976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1216..1235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1563..1584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1719..1739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1926..2037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2155..2295
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2155..2284
                     /label=AAV2 ITR
     rep_origin      2370..2825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2457..2476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2667..2688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2842..2861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2961..2983
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3021..3039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3107..3211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3212..3280,3281..4072)
                     /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(3430..3449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4243..4831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4732..4751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttccac ttatacccca tcagcatctc
      181 tatcattgtc taagacattt ttaatttgaa ataaatgggg aaactcatta tcatggcctc
      241 aaggaaatac agaaaccaag gattgtcctt ttagtgcaca ggtgtcttct aaattcacat
      301 attctggatt taacaaggaa aataaaatct gacataggct gaatttgaaa atggtaagga
      361 tatttttaac taaacaaaaa aaataggtaa tagaacattt catattgaac tcccaggata
      421 gcatttatca taatccacca gttgaattat tcatgcgctg ataaataacc ttagcctccg
      481 gcacagtaat cctctcactg aataatcacc agggctccct gcccgtggtg ttttgcttta
      541 caacataaat tttaaaatta ttctgtgttg gcatacatat gtgaaagctc attaacagat
      601 actggcattc tgattggttt ggcactaaat ttaaactgtt ttggaaagaa ctgaaacact
      661 gaaggatatg ccattctgtt tctttatacc ttatcgatgt ttccttaaca atcggttgag
      721 atttatgcat cttttgatag attttatctg agatagctga tatgcattgt gatgagttcc
      781 tttttaacgc tgtaatgttt cattggccct gagctcgggc tgggcataaa agtcagggca
      841 gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg
      901 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1021 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     1081 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
     1141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     1381 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1501 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1621 tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat
     1681 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1741 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1801 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     1861 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc
     1921 ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1981 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2041 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2101 ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac
     2161 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     2221 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     2281 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2341 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     2401 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2461 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2521 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2581 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2641 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2701 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2761 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     2821 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2881 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2941 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3001 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3061 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3181 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3241 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3301 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3361 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3421 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3481 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3541 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3601 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3661 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3721 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3781 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3841 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3901 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3961 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4021 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4081 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4141 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4201 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4261 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4321 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4381 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4441 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4501 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4561 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4621 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4681 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4741 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4801 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4861 cctggccttt tgctggcctt ttgctcacat gt
//
