LOCUS       sequence-450566-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0774m 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 4852)
  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 4852)
  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: AiP12606 - pAAV-AiE0774m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4852
                     /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      864..873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(870..872,873..875,876..1586)
                     /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(915..936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1176..1195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1523..1544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1679..1699)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1886..1997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2115..2255
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2115..2244
                     /label=AAV2 ITR
     rep_origin      2330..2785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2417..2436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2627..2648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2802..2821)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2921..2943
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2981..2999)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3067..3171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3172..3240,3241..4032)
                     /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(3390..3409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4203..4791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4692..4711
                     /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 gcgttccaga tgcttgtctt ctctctctct ctctctctct
      181 ctctctctct ctctctctgt atgtgtgtgt gtgtgtgtgt gtgtatgtgt gtcttccccc
      241 ctctttttca ccccaccccg acatctacaa caaaaatttt aaccatatta tggagtggtc
      301 atattggcag tttttttact gcagcccttc acatgcactg acattctaga cagcaagtgt
      361 tctatgattt tcaacaggac acttggatct agccaactca gttttcaagc ctgcccccga
      421 gttgtagatc ctgtcttcac attctaaact aatctcacac cactttgtgc caactccact
      481 ctggtaaaga ccatgcactc gagttagtgc aagacatttt gttctggcct cggggtgtgc
      541 tgccgcctcg tcaggatggg aggtggaggc gagtccagtg aaggtcactg gcatcacaag
      601 agtgagggac agatatgttc ccgagccttc agattcaagt cctgcacttg caaaaaagac
      661 ttctgccttt tctatttgac ccaactagct attgcattat tgagccagac agtggctcaa
      721 attaaatgtc acaggagcac accctaacaa aaagaagagg ccagcatgct gagctcgggc
      781 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc
      841 tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
     1021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
     1081 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1321 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1381 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1441 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1501 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1561 ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat
     1621 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1681 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1741 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     1801 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     1861 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1921 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1981 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2041 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     2101 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2161 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2221 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2281 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2341 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2401 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2461 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2521 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2581 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2641 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2701 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2761 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     2821 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2881 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2941 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3001 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3061 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3121 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3181 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3301 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3361 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3421 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3721 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3781 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4021 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4081 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4141 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4321 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4381 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4441 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4501 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4561 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4621 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4681 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4741 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4801 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
