LOCUS       sequence-466302-        4981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2382m 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 4981)
  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 4981)
  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: AiP1765 - pAAV-AiE2382m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      385..394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(391..393,394..396,397..1107)
                     /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(436..457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(697..716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1044..1065
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1200..1220)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1407..1518
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1636..1776
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1636..1765
                     /label=AAV2 ITR
     rep_origin      1851..2306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1938..1957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2148..2169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2323..2342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2442..2464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2502..2520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2588..2692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2693..2761,2762..3553)
                     /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(2911..2930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3724..4312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4213..4232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4374..4503
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 caatttctca gctaacaaat ggtcactaag gaaagtacca gagagcagcc ctgattctgt
       61 ggatgtcact gacaggaacc ttcaggatgg gcaaccccaa gatggcctgc tgcctgctca
      121 cctcactctc tcccatatac caccttttac aactctttgt gacaatttaa ttaagacgca
      181 acccttggga tttggatgca ttgaatgtaa gtggcaagat cagaacaatc tccttcctgt
      241 ccccttctta cgaagcaatt caaagcttgt ggtttctatt caatttgttc ggagctcggg
      301 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      361 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      421 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      481 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
      541 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
      601 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      661 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      721 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
      781 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
      841 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
      901 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
      961 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1021 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1081 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1141 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1201 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1261 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1321 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1381 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1441 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1501 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1561 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     1621 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     1681 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     1741 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     1801 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     1861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1921 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1981 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2041 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2161 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2221 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2281 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2341 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2401 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2461 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2521 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2581 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2641 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2701 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2761 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2821 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2881 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2941 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3001 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3061 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3121 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3181 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3241 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3601 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3661 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3721 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3781 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3841 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3901 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3961 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4021 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4081 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4141 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4201 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4261 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4321 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     4381 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     4441 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     4501 cctgcggccg cacgcgttgt aagagtaata atcgcaatct cagaaagatt cgtagatttt
     4561 tatatagcta cacaaagtta tatattcaca gatggcataa atatagaagg gaaagtatgt
     4621 aggtggccac ggggactcat aggagagaaa agggaaaggg gggagggatt atgaggaaga
     4681 gtaggcagga cacacattat attcttgtat gaaaacttta aaaaacaagc tttaaaaata
     4741 aaccctaatt atcaagagaa tatttcacac agccctccac agcaatttct ccaggtaagg
     4801 aagaagatag tgatgcttcc atcaacactg tgagatgtgt tcagatgcct ccaggtttga
     4861 aatggggata agaaaaaggc tacgttcagc tgtgtaaatc acgcaggaag tggctgctgg
     4921 gaaggaagga aatggaatcc cattttcctt cttgttttgc tcctgaaact tctgcactac
     4981 a
//
