LOCUS       sequence-466234-        5748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2066m 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 5748)
  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 5748)
  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: AiP1281 - pAAV-AiE2066m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..54
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   196..325
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1955..1964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1961..1963,1964..1966,1967..2677)
                     /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(2006..2027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2267..2286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2614..2635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2770..2790)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2977..3088
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3206..3346
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3206..3335
                     /label=AAV2 ITR
     rep_origin      3421..3876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3508..3527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3718..3739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3893..3912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4012..4034
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4072..4090)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4158..4262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4263..4331,4332..5123)
                     /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(4481..4500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5294..5748,1..134)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
       61 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      181 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      241 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      301 gccaactcca tcactagggg ttcctgcggc cgcacgcgtc agtggtgttg gttaaaggtc
      361 aacagatgag gttcagagca catcaagtgt atgggaagcc accgatacct ttgaagctgg
      421 agaggccatg atctgatgat gttttaaggc tgtcttctga aagcctagct ggagaagaga
      481 ttttacagcc accatgtaac ctatgaagaa accaagcagg aacttatggc aatgtgtccc
      541 tgggacagga aggcaccaga gggagaggag tggggctgga agcagaccat tgcaagataa
      601 tgaagagtct tcctgaagat tgcagagctt ctgatacgaa aggctcaacc attccagatc
      661 acccattttg catccagttc tgatgttcca ccagtgaact tgacttcatt caggtactgg
      721 gtttgttccc aaccaaatga accacgagca tagatgggaa ggggaacaca atgaaggatg
      781 atgttggagc cggtgaagat gagaaggccc tatggaaact gtcaactgcc tatgcccaga
      841 acttcttcac agtatggagg tttatattac ctaatatatg ccccaacttt aattaataat
      901 gtccaatagg cactatagat tatatactct gtatatggaa caagtatatt ttcaattcaa
      961 ttaaaatact taagattaaa ccaatgtcat ctcaaattga aactcagtct ccctacttcc
     1021 ttgtttccat tttattgcca tgttttcctc cctaccatac aactagcatt ttttttaact
     1081 tgctgtcaga ctttccccat tcagcagtga ccgaaacaaa gggaaaaaat ggtacagagg
     1141 gaaaatcagc agttctgttc aataatggcc aattgtttcc ctgacagcat ccagacaaat
     1201 cgtctaaatc acaaatgcat ttgtgaccct cattttggaa gggcttctct aatcttaggg
     1261 attggaattg aacttcaaaa atgggtctca aggtctttgg gggcccctca atttgctgtt
     1321 ccaacaccct agtggctgga tggctaccat cctttttcat tcgcttaata aaaataacaa
     1381 gaaaaactat aatagccaaa tttcttctct caaatgccaa tgtagagagc aatcattccc
     1441 aaactctgag gcagtaaaga ggtgactttg aacaatgcct ttattttttc ctcatttatt
     1501 tctcaatttt tatctccaga ttagatatgt gcctatagga agggaagcag tgctgggaag
     1561 tgcagagtag tttgatgtca cacagcagat taagcaaaga atcatgtata atcaccactg
     1621 aatgcccatg tgcatccttc catcttgtgg ctttgtgtgg gagagtcaaa ggtggtctgg
     1681 atttcaaatg aacggaaaaa tcaccctgtg actggataga ggttccagga cacactctct
     1741 ctccctctct ctttccctct ctccctctct tttaggaaac accaaaagat gtagctctca
     1801 ttggataaaa taaaatatgg gcaaaaacat catggatact ggcatgcatt agtattaagt
     1861 cgagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     1921 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     1981 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2041 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     2101 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     2161 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2221 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2281 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2341 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2401 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     2461 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     2521 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     2581 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2641 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     2701 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2761 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2821 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     2881 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2941 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     3001 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3061 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3121 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     3181 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3241 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3301 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3361 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3421 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3481 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3541 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3601 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3661 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3721 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3781 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3841 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3901 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3961 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4021 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4081 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4261 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4321 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4381 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4441 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4501 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4561 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4621 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4681 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4741 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4801 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4861 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4921 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4981 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5041 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5101 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5161 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5221 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5401 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5461 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcaggg
//
