LOCUS       sequence-450575-        5001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0772m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L6_IT_Car3 neurons (note CLA 
            neurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5001)
  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 5001)
  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: AiP12612 - pAAV-AiE0772m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5001
                     /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      1013..1022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1019..1021,1022..1024,1025..1735)
                     /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(1064..1085)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1325..1344)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1672..1693
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1828..1848)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2035..2146
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2264..2404
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2264..2393
                     /label=AAV2 ITR
     rep_origin      2479..2934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2566..2585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2776..2797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2951..2970)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3070..3092
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3130..3148)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3216..3320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3321..3389,3390..4181)
                     /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(3539..3558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4352..4940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4841..4860
                     /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 gcgtaggagg aaatgtgctg tggtttaact tttggaataa
      181 aaatagtgaa tttttaatga gcaggtggga atgtccttgg gttcctgagt taggatggtg
      241 gagcaaggaa agaaaaggtc tctcttcctt ttccctgtgt gctgagtaga gacaggattg
      301 cccagaacac caggaaccta ggttgtactg acaaaagtgg aaaacagccg caaaaccgct
      361 agggaaagca gctgtcccct tgcccagtct cctgccccac tcctgcctct gcctataaga
      421 gtttggcaca gttggagatt tgtatacaca aatgcatgcg tggaaatact gcgctctgat
      481 gacattcctc agggggcgac ctttgatgat gaattttctt gcccttttta ggggaggaga
      541 agaggaggcg ggtggtactt cacaagacta aatttactca acttttttgg gtctaaattt
      601 ctttaggaaa aaaaatgtca aatgattgta gatcccaaac tgtctttgtt ggcatttgaa
      661 tggctctgtg agtccagtgt ggggagcaag tgaaaagaaa agccacctct cccctcgctg
      721 gcgtggtatt gaagctgaag ccctggacca cacagtgtct tggggacctt catgatttat
      781 atattgtatg tgatcttttc ttaaacattg ttaacctgtg ccctttacag acaatgccgg
      841 gtcagaaaat gtcaggggaa cttcggactg ctttcataat tgtgtaaaga aaattgttgt
      901 cttcagaatc cagagcccag agctcgggct gggcataaaa gtcagggcag agccatctat
      961 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
     1021 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1081 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1141 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1201 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1261 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1321 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1381 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1441 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1501 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1561 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1621 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1681 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt
     1741 cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg
     1801 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1861 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1921 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1981 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     2041 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2101 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2161 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2221 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     2281 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2341 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2401 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2461 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2521 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     2821 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     2881 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     2941 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     3001 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3061 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3121 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3181 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3241 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3301 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3421 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3481 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3541 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3601 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3721 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3841 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3901 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4141 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4981 gctggccttt tgctcacatg t
//
