LOCUS       sequence-433040-        4779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2369m 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 4779)
  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 4779)
  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: AiP1694 - pAAV-AiE2369m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..91
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(129..147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        215..319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(320..388,389..1180)
                     /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(538..557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1351..1939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1840..1859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2001..2130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2791..2800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2797..2799,2800..2802,2803..3513)
                     /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(2842..2863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3103..3122)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3450..3471
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3606..3626)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3813..3924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4042..4182
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4042..4171
                     /label=AAV2 ITR
     rep_origin      4257..4712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4344..4363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4554..4575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4729..4748)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
       61 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      121 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      181 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      241 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      301 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      361 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      421 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      481 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      541 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      601 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      661 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      721 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      781 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      841 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      901 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      961 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1021 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1081 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1141 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1201 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1261 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1321 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1381 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1441 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1501 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1561 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1621 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1681 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1741 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1801 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1861 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1921 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1981 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2041 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2101 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgttgtaag agtaataatc
     2161 gttggattgg gtcccaggct gtatacaaaa aatccttgca gacagaagag ctttgtctgg
     2221 gccccctgtg gaatgtagct ggattctaaa gtggctgcag tgccctatga taagcagtat
     2281 gtgtcccagc actagtggct atgattgtgc tctctggagc taacagcatc caaagcaatt
     2341 gatgccagat aaacagtagc tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg acagtacatt
     2401 ttgaaaagac aggacatttt gtctctcttc cttgttccaa ctgtccttag ctcttcctgg
     2461 acacaatgag tgattagatt aagaggaaga tggtcctttg ctctcaggac accaatgagc
     2521 ccactcttag cacggagttg gactccaaat actacagtac tagccaggag cttttgtaca
     2581 acaccaggat accttggatg ctaaattcag aatgtcttac atggtattag agaacaatga
     2641 agccaggcac gggacacatg gcaaccacag ggggaggggt ggagcttggg agatgcggag
     2701 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
     2761 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
     2821 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2881 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     2941 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     3001 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3061 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3121 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3181 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3241 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     3301 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     3361 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     3421 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3481 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     3541 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3601 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3661 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     3721 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3781 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     3841 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3901 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3961 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     4021 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4081 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4141 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4201 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4261 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4321 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4381 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4441 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4501 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4561 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4621 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4681 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     4741 tgatgccgca tagttaagcc agccccgaca cccgccaac
//
