LOCUS       sequence-473571-        5420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0924m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5420)
  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 5420)
  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: AiP14096 -
            pAAV-AiE0924m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5420
                     /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      773..782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(779..781,782..784,785..1495)
                     /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(1085..1104)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1432..1453
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1505..1561
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1574..1639
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1670..2047
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2134..2154)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2341..2452
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2570..2710
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2570..2699
                     /label=AAV2 ITR
     misc_feature    2717..2744
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2745..2772
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2841..3296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2928..2947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3138..3159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3313..3332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3432..3454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3492..3510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3578..3682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3683..3751,3752..4543)
                     /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(3901..3920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4714..5302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5203..5222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5359..5386
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5387..5414
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttcagag gccagtgtgt atttagggta gggcagtaca
      181 ttaaagttct ggtttgtgtg ctgtcaccct atcctagaaa ctggtttgag agttttagtt
      241 cacatgcaaa atgaaggtca aagtcaatac caaggagtct aaaatcaacg tgaagtgata
      301 cagagatgtg cacagtttgt ttgttacgaa catcactgcc tcctacagaa tgaacttatc
      361 atcatgtgtc acaatcatcc catgcatttt ccagctactt ccagtgagct ccaaatcttt
      421 acacattttg aggttgaagt aaaagagcaa tgtgcacagt gagatagtca cagtttcttt
      481 acccagtctt gggtgattca gattcagtca ggccaacagg gaggactttc ctgccaggag
      541 gggggcccga gcaagaacag gagcttagac cctatgatgc ttagaaagaa cccaggaaac
      601 tgcctatgct gtctgttgat tatgccgtga ttgctgcagg aaatgcagta gtttatgaaa
      661 cagtagaagc taagcatgtg agctcgggct gggcataaaa gtcagggcag agccatctat
      721 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
      781 ggtctccaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      841 cgatgtcaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      901 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      961 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1021 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1081 caaagacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1141 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1201 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1261 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1321 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1381 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1441 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggctc
     1501 tggtgctacc aacttctcac tgttgaaaca ggcaggggat gtagaggaga atccagggcc
     1561 tggtgctagt ggagactaca aagaccatga cggagattat aaagatcatg acatcgatta
     1621 caaggatgac gatgacaagt ccggactcag atctggaggc tccggaggcc cagagccagc
     1681 gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg cgcagaagaa
     1741 aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg tgtataaggt
     1801 tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca tcatgaactc
     1861 gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg cgcattacaa
     1921 caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc tgctgcctgg
     1981 ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt acaccagcgc
     2041 taagtaatga ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     2101 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2161 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2221 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     2281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     2341 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2401 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2461 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2521 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     2581 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2641 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2701 tgcctgcagg ggcgcctatc agcacacaat tgcccattat acgcgcgtat aatggactat
     2761 tgtgtgctga tatgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2821 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3181 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3241 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3301 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3361 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3421 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3481 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3541 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3601 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3661 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3721 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3781 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3841 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3901 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3961 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4021 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4081 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4141 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4201 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4261 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4321 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4381 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4441 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4501 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4561 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4621 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4681 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4741 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4801 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4861 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4921 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4981 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5041 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5101 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5161 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5221 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5281 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5341 ttgctggcct tttgctcata tcagcacaca attgcccatt atacgcgcgt ataatggact
     5401 attgtgtgct gataacatgt
//
